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  • Saccharomyces cerevisiae  (759)
  • Angiosperms
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  • MDPI - Multidisciplinary Digital Publishing Institute  (25)
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  • 1
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-01-08
    Description: Wine has a complex matrix with many volatile compounds present, which evolves over time. These volatile compounds are important to wine quality as they contribute to the aroma and varietal characteristics of wine. Recent development in the analysis of volatile compounds in wine has greatly improved our understanding of the complexity of wine aroma. Analytical methods used for wine aroma fingerprinting have shown potential in determining the origin and quality of wine. Thus, research on volatile compounds responsible for wine aroma and their correlation with wine provenance and wine quality have increasingly attracted great interest from researchers and winegrowers. This Special Issue presents the latest research regarding wine aroma compounds, including, but not limited to, the topics on the characterization of aroma compounds in grapes and wine, factors influencing the production of aroma compounds in wine during fermentation and maturation, and analytical methods for wine aroma analysis.
    Keywords: marselan wine ; aroma compounds ; indigenous yeast strains ; Saccharomyces ; non-Saccharomyces ; icewine ; Vidal ; yeast ; sensory analysis ; amino acid ; fruity ester ; wine aroma ; nitrogen management ; Pearson correlation analysis ; carbon metabolism ; Vitis davidii Foёx ; spend coffee grounds ; fermentation ; sensory property ; volatile profile ; yeast protein hydrolysate ; nitrogen supplementation ; volatile compounds ; wine higher alcohols ; wine esters ; monoterpenes ; triangle test ; check-all-that-apply ; correspondence analysis ; Cochran’s Q-test ; nutrients ; central composite design ; Saccharomyces cerevisiae ; wine ; strain effect ; aromas ; non-Saccharomyces yeasts ; ethanol tolerance ; ultraviolet irradiation ; diethyl sulfate mutagenesis ; vineyard mechanization ; phenolics ; sensory properties ; anthocyanins ; bentonite ; cold soaking ; colour ; pathogenesis-related proteins ; Pinot noir ; tannin ; antioxidants ; glutathione ; glutathione-enriched inactivated dry yeasts ; methoxypyrazines ; oxidation ; Sauvignon Blanc ; thiols ; aroma profile ; grape pomace ; model juice ; bic Book Industry Communication::G Reference, information & interdisciplinary subjects::GP Research & information: general ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences ; bic Book Industry Communication::T Technology, engineering, agriculture
    Language: English
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  • 2
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2023-04-05
    Description: The eradication of vector-borne diseases is threatened by the limited range of available insecticides, leading, inevitably, to the development of resistance. This is particularly concerning for malaria control, which relies heavily on insecticide-treated nets (ITNs) and indoor residual sprays (IRS). New chemistries are being developed, and innovative deployment of insecticides may play a role in overcoming resistance, either through new types of tools or new means of distribution. A variety of novel product types and vector control strategies are under development and evaluation, which is to be celebrated, but a strong evidence base is needed to guide effective operational deployment decisions. Novel approaches should be supported by robust data collected using appropriate and validated methods to monitor efficacy, durability, and any emerging resistance. This reprint presents original research into developing and characterizing new vector control products, as well as understanding and monitoring insecticide resistance. Review articles explore the impact of insecticide resistance and offer guidance on insecticide choice in the face of pyrethroid resistance. Consensus methodologies are presented, in the form of standard operating procedures (SOPs) designed to be adopted and used to generate reproducible data that can be compared and interpreted across and between studies. It is hoped that this collection of articles offers inspiration and guidance on how consistent data can be generated to inform more effective development, evaluation, and use of new and existing vector control tools.
    Keywords: prallethrin ; insecticide ; spatial treatment ; mosquito fitness ; protection ; pyrethroids ; Aedes albopictus ; Culex pipiens ; life tables ; mosquito ; bite-proof garment ; model ; textile ; non-insecticidal ; physical barrier ; insecticide selection ; out-crossing ; strain authentication ; laboratory screening ; pyrethroid ; pyrethroid resistance ; insecticide resistance ; insecticide resistance management ; vector control ; malaria ; malaria control ; Anopheles ; host-seeking behavior ; insecticide exposure ; pathogen transmission ; Aedes aegypti ; Anopheles gambiae ; ATSB ; Culex quinquefasciatus ; Iroquois ; RNAi ; Saccharomyces cerevisiae ; yeast ; Anopheles mosquito ; fertility ; ovary development ; pyriproxyfen (PPF) ; side-effects ; machine learning ; image classification ; automated identification ; convolutional neural network ; insecticide-treated net (ITN) ; PBO ITN ; synergist ITN ; dual-AI ITN ; insecticide resistance management (IRM) ; method validation ; durability monitoring ; bioinsecticide ; disease transmission ; insecticide-resistance ; mosquito-borne disease ; mosquito control ; natural compounds ; phytochemical ; malaria vector ; insecticide treated nets ; cytochrome P450s ; kdr ; cuticular resistance ; deltamethrin ; imidacloprid ; bifenthrin ; β-cyfluthrin ; etofenprox ; α-cypermethrin ; λ-cyhalothrin ; thiacloprid ; mosquitoes ; Attractive Toxic Sugar Bait (ATSB) ; Attractive Targeted Sugar Bait (ATSB) ; diagnostic bioassay ; resistance monitoring ; insecticide-treated nets (ITN) ; strain characterisation ; method development ; product evaluation ; quality control (QC) ; dual active ingredients (dual-AI) ; bioefficacy ; IRS ; application technology ; broflanilide ; clothianidin ; pirimiphos-methyl ; WHO tube ; WHO tunnel test ; ITNs ; interceptor ; interceptor G2 ; membrane ; human arm ; rabbit ; bioassay ; bio-efficacy ; n/a ; bic Book Industry Communication::G Reference, information & interdisciplinary subjects::GP Research & information: general ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences::PSB Biochemistry
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  • 3
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2023-08-08
    Description: The externalization of animals’ genetic potential is determined by a number of external factors, of which feeding plays a major role. Animals’ nourishment is one of the most important levers to ensure the efficiency of animal production due to both the high share of feed costs in the total cost of products and the influence this has on the growth, reproduction, and health of animals as well as the quality of products obtained from these. This field is one of the most dynamic in the field of husbandry sciences due to the takeover and permanent use of numerous results obtained from research on energy metabolism and nutrients related to the composition of feed and its influence on animal products. This is also due to the great advances in genetics, which create new types of animals with increasing productive potential, but also with different food requirements. This Special Issue collated innovative papers on animal nutrition, physiology, chemistry, biochemistry, genetics, reproduction, and breeding technologies. The articles covered a wide range of topics related to feed quality, the influence of food on the production level, the quality of production, and also on animals’ health.
    Keywords: carcass yield ; commercial cuts ; low cost ; neutral detergent fiber ; non-fiber carbohydrate ; Yucca schidigera ; antimicrobial ; secondary metabolites ; sustainability ; pollution ; production ; food animals ; Ajuga iva ; chemical composition ; nutritive value ; unconventional feeds ; phenolic ; growing conditions ; dairy buffaloes ; farming environment ; reproductive and productive performances ; feeding trial ; mozzarella cheese ; sensory properties ; alternative feed ; degradability ; fractions ; ram ; sperm quality ; Saccharomyces cerevisiae ; apparent digestibility ; honey ; quality ; phenolic content ; flavonoid content ; Pearson’s correlation ; female camels ; milk ; minerals ; heavy metals ; winter ; total mixed ration ; paddlefish ; meat quality ; fatty acids ; biological value ; body condition score ; ewes ; reproductive traits ; flushing ; animal production ; genetic diversity ; grey cattle ; mitochondrial DNA ; Podolian cattle ; European catfish ; somatometry ; corporal indice ; flesh yield ; nutritional quality ; lactation ; manganese ; reproductive performance ; sows ; AP monitoring ; IoT ; AP estimation ; decision support ; livestock farming ; polycyclic aromatic hydrocarbons ; meat ; chicken ; duck ; turkey ; phenols ; flavonoids ; FTIR ; rearing system ; birds’ welfare condition ; biochemical analysis ; productive parameters ; food and feed safety ; yeasts and molds ; Salmonella spp. ; Escherichia coli ; Clostridium perfringens ; rabbit ; hare ; lipid health indices ; water-holding capacity ; cooking loss ; egg weight ; shell weight ; fractional reduction ; deletion method ; reproduction ; n/a ; bic Book Industry Communication::G Reference, information & interdisciplinary subjects::GP Research & information: general ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences
    Language: English
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  • 4
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2023-04-05
    Description: Adaptation to external changes is necessary for all cells to survive and thrive in diverse environments. Key to these responses are the MAPK-mediated signaling pathways, intracellular communication routes that sense stimuli at the cell surface, and are ubiquitous in all eukaryotic organisms. In the case of fungi, MAPKs mediate essential processes, such as adaptation to environmental stresses, morphology regulation, or developmental processes. First studied in the early nineties in Saccharomyces cerevisiae, the fungal cell wall integrity (CWI) pathway has proven to be a central MAPK-mediated signaling cascade conserved in the fungal kingdom. Cells need to sense cell wall-perturbing conditions and mount the appropriate salvage response. Understanding this CWI pathway-mediated compensatory mechanism is key for the development of cell wall-targeted antifungal therapies. Moreover, its functional roles go beyond the maintenance of this essential structure, reaching many other physiological aspects that have major implications in development or virulence.In this Special Issue, expert researchers in this relevant subject have contributed with seven reviews and eleven original articles to advance our understanding of the CWI pathway by covering different structural, regulatory, and functional aspects in distinct yeasts and filamentous fungi.
    Keywords: Wsc1 ; membrane sensor ; SMALP ; detergent-free extraction ; fluorescence correlation spectroscopy ; transmission electron microscopy ; 3D reconstruction ; fission yeast ; MAPK ; cell integrity pathway ; S. japonicus ; S. pombe ; protein kinase C ; Pmk1 ; dimorphism ; hyphae ; yeast ; cell wall integrity ; phosphorylation ; azoles ; clotrimazole ; cytokinesis ; actomyosin ring ; septum ; cell integrity ; fungi ; cell wall ; cell wall proteins ; signaling pathways ; stress tolerance ; mannoprotein ; budding yeast ; morphology ; CalMorph ; cell wall integrity (CWI) pathway ; PKC ; GTPases ; MAP kinase ; morphogenesis ; virulence ; pathogenesis ; Hrr25 ; Mec1 ; Tel1 ; Pkc1 ; hydroxyurea ; UV irradiation ; cell wall integrity (CWI) ; Mtl1 ; autophagy ; glucose ; mitophagy ; Saccharomyces cerevisiae ; histidine kinase ; Paracoccidioides ; paracoccidioidomycosis ; cell cycle ; Slt2 ; checkpoint ; DNA damage ; conjugation ; ploidy ; lysis ; Cell Integrity Pathway ; stress ; CWI pathway ; UPR ; glucosamine ; tunicamycin ; N-glycosylation ; cell wall integrity pathway ; MAPK substrate ; kinase assay ; fungal cell wall ; cysteine-rich domain ; PAN domain ; aromatic clusters ; filamentous fungi ; signaling pathway ; surface sensor ; mitogen-activated protein kinase ; plant pathogen ; application ; fungicide ; drug target ; culture ; productivity ; stress response ; screening ; transcription ; essential genes ; n/a ; bic Book Industry Communication::G Reference, information & interdisciplinary subjects::GP Research & information: general ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences
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  • 5
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-03-28
    Description: Explore a decade of groundbreaking research in "10th Anniversary of Cells—Advances in Plant, Algae, and Fungi Cell Biology." This reprint offers a comprehensive journey into the realms of plant, algae, and fungi cell biology. Delve into the world of genomics, cellular defense mechanisms, mycorrhizal fungi, and the physiology of extremophile algae. A celebration of scientific excellence, this reprint is a valuable resource for researchers, educators, and enthusiasts passionate about these fascinating domains. Join us in commemorating a decade of discovery and advancement in cellular biology.
    Keywords: membrane proteins ; overproduction ; production platform ; protein purification ; Saccharomyces cerevisiae ; solute carrier 39 ; SLC39 ; family ; yeast ; zinc ; zinc transporters ; ZIPs ; Agave americana ; crassulacean acid metabolism ; genetic engineering ; Nicotiana sylvestris ; phosphoenolpyruvate carboxylase ; photosynthesis ; drought tolerance ; salt tolerance ; microalgae ; Chlamydomonas reinhardtii ; starch ; supraoptimal temperature ; cell cycle ; pilot-scale production ; DNA methylation ; Fusarium graminearum ; in vitro subcultures ; virulence reduction ; ddRAD-MCSeEd ; virulence genes ; 13C ; 14C ; aldol ; Calvin-Benson cycle ; light respiration ; isotope labeling ; cytokinin ; endocytosis ; cytoskeleton ; actin ; plant immunity ; induced resistance ; Parachlorella kessleri ; supra-optimal temperature ; energy reserves ; growth processes ; reproduction events ; deuterium ; deuterated starch ; deuterated lipid ; soft scale insects ; Ophiocordyceps ; symbiosis ; transovarial transmission ; Verticillium wilt ; Glomus viscosum Nicolson ; arbuscular mycorrhizal fungi ; oxidative stress ; antioxidant systems ; defense ability ; ABI5 ; ABF ; AREB ; abiotic stress response ; abscisic acid ; phytohormone crosstalk ; salinity stress ; chloroplast ; plastid ; osmolytes ; osmotic adjustment ; reactive oxygen species ; herbivory ; membrane potential ; ion channel ; Arthrospira ; haloalkalotolerant cyanobacteria ; metagenomics ; phylogenomics ; fatty acid ; enveloped virus ; Ebola virus ; HIV ; herpes simplex virus ; human cytomegalovirus ; influenza virus ; MERS-CoV ; SARS-CoV-2 ; N-glycosite ; O-glycosite ; high-mannose glycan ; complex N-glycans ; Vicieae man-specific lectin ; T/Tn-specific lectin ; specific interaction ; n/a ; thema EDItEUR::G Reference, Information and Interdisciplinary subjects::GP Research and information: general ; thema EDItEUR::P Mathematics and Science::PS Biology, life sciences
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  • 6
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-04-11
    Description: This Special Issue, “Biotechnology Applications of Microalgae”, is focused on the latest novel advances related to the production of different bioactive compounds from microalgae and their biotechnological use.
    Keywords: enzymatic activity ; fluid dynamics ; microalgae ; oxidative stress ; static magnetic fields ; violaxanthin ; reactive oxygen species ; ascorbic acid ; glutathione ; tocopherols ; phenolic compounds ; carotenoids ; thraustochytrids ; antioxidants ; saturated fatty acids ; polyunsaturated fatty acids ; transcriptomics ; sustainability ; industrial valorization ; carbon dioxide fixation ; biological activities ; phytosterol ; Saccharomyces cerevisiae ; Phaeodactylum tricornutum ; Sparus aurata ; β-glucans ; pulse feeding ; immune tolerance ; salt stress ; seawater cultivation ; Internet of Things ; proteomics ; blue light ; astaxanthin ; fatty acid ; heme ; cell wall ; salicylic acid ; fucoxanthin ; green consumption ; food consumption ; amino acids ; carbohydrates ; radical scavenging activity (RSA) ; RP-HPLC ; Chromochloris zofingiensis ; lutein ; CO2 aeration ; cGMP-dependent kinase ; biodiesel ; microalgal biotechnology ; natural antioxidants ; Yarrowia lipolytica ; Chlorella vulgaris ; growth ; fatty acids ; Spirulina ; healthcare ; space missions ; medicine applications ; microgravity effects ; humic substances ; microalgae cultivation ; hormetic effects ; increased nutrient availability ; improved protection against abiotic stress ; higher accumulation of bioactive ingredients ; enhanced microalgal productivity ; Dunaliella salina ; chlorpropham ; herbicide ; phytoene ; Nannochloropsis ; mixotrophy ; photobioreactors ; CHN analysis ; metabolomics ; bioassay ; cell death pathway ; autophagy ; antitumoral activity ; thema EDItEUR::T Technology, Engineering, Agriculture, Industrial processes::TB Technology: general issues ; thema EDItEUR::T Technology, Engineering, Agriculture, Industrial processes::TC Biochemical engineering::TCB Biotechnology
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  • 7
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-03-28
    Description: This reprint presents recent developments in the field of biological liquid–liquid phase separation (LLPS, also known as biomolecular condensation). LLPS and related biogenesis of various membraneless organelles (MLOs) and biomolecular condensates (BMCs) represent fundamental molecular mechanisms governing the spatio-temporal organization of the intracellular space. In fact, MLOs and BMCs, being liquid droplets, represent specific compartments within a cell that are not enclosed by a lipid membrane. Most biological LLPS processes are reversible, and many MLOs/BMCs exist transiently; they rapidly emerge when conditions are changed and rapidly disintegrate as soon as the original conditions are restored, thereby showing a characteristic “now you see me, now you don’t” behavior. Numerous MLOs/BMCs are found inside eukaryotic cells, where they exist as liquid droplets (or cellular bodies, puncta, etc.) in the cytoplasm, nucleoplasm, mitochondrial matrix, and stroma of chloroplasts. Furthermore, MLOs/BMCs are commonly observed in Archaea, bacteria, and, likely, viruses. MLOs/BMCs have numerous crucial functions, and their biogenesis is known to be controlled by various external factors and environmental cues, such as changes in temperature, pH, and ionic strength of the solution. All of these have garnered the close attention of many researchers to biological LLPS, MLOs, and BMCs.
    Keywords: Alzheimer’s disease ; amyloid aggregation ; lipid bilayer ; cholesterol ; time-lapse AFM imaging ; molecular dynamics ; liquid–liquid phase separation (LLPS) ; membraneless organelles ; phase-separated condensates ; human diseases ; liquid–liquid phase separation ; intrinsically disordered proteins ; proteins with low complexity ; P-body ; Nst1 ; polyampholyte domain ; aggregation-prone domain ; Saccharomyces cerevisiae ; membrane-less organelle ; nuclear speckle ; nucleolus ; phase separation ; chromatin organization ; nuclear condensate ; intrinsically disordered region ; transcription ; DNA damage repair ; super-enhancer ; quantitative imaging ; CTP synthase ; cytoophidium ; fluorescence recovery after photobleaching (FRAP) ; stimulated emission depletion (STED) ; Drosophila ; epithelium ; follicle cell ; ingression ; paramyxoviruses ; Hendra virus ; amyloid-like fibrils ; Taylor Dispersion Analysis (TDA) ; negative staining Transmission Electron Microscopy (ns-TEM) ; Polyethylene glycol (PEG) precipitation assays ; Congo Red ; Small-Angle X-ray Scattering (SAXS) ; actin ; actin polymerization ; actin-binding proteins ; coacervate ; membrane ; signaling proteins ; n/a ; thema EDItEUR::G Reference, Information and Interdisciplinary subjects::GP Research and information: general ; thema EDItEUR::P Mathematics and Science::PS Biology, life sciences
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  • 8
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2023-04-05
    Description: Tryptophan is a rate-limiting essential amino acid and a unique building block of peptides and proteins. This largest amino acid serves as the precursor for the important endogenous indoleamines serotonin, N-acetylserotonin, and melatonin that act as neurotransmitters, neuromodulators, and neurohormones. Kynurenic acid is the most potent endogenous antiexitotoxic agent. Other highly relevant pathways of tryptophan are the reversible transamination to indole-3-pyruvate with formation related indolic acids that act as potent antioxidant agents. Tryptophan metabolites, such as melatonin, and structurally related agents, such as indole-3-propionic acid, act as potent catalytic antioxidants and bioenergetic agents that facilitate regeneration and protection against stress and aging. Several indole compounds act as uremic toxins since these agents can induce radical formation that is associated with enhanced oxidative stress and damage. The exploration of the effects of these protective and toxic tryptophan derived agents has revealed important molecular mechanisms and mediators of adaptation and aging. Research on tryptophan in nutrition and health can facilitate the development of new approaches to extend human health and life span. Amino acids are the building blocks of life that enable repair, as well as recycling and regeneration. Research on nutrients like amino acids, such as tryptophan and its metabolites, as well as peptides and proteins, or extracts containing this molecular metabolism modifiers can improve health. Research into the indololome is a new emerging and rapidly growing field of utmost relevance to science and society.
    Keywords: tryptophan ; kynurenine ; kynurenic acid ; FICZ ; AhR ; melanoma ; proliferation ; cell death ; aryl hydrocarbon receptor ; chronic kidney disease ; developmental origins of health and disease (DOHaD) ; hypertension ; indole ; melatonin ; serotonin ; uremic toxin ; virus ; immunity ; codon ; depression ; chronic mild stress ; oxidative stress ; tryptophan catabolites pathway ; methylation ; expression ; escitalopram ; 5-hydroxytryptophan ; natural sources ; microbial production ; biosynthetic pathways ; physiological effects ; animal ; human ; kynurenine pathway ; MEL biosynthesis ; Saccharomyces cerevisiae ; yeast ; tryptophan extraction ; LC-MS/MS ; soybean ; skin ; atopic dermatitis ; psoriasis ; severe acute respiratory syndrome ; SARS-CoV-2 ; COVID-19 ; malignant melanoma ; urine ; autofluorescence ; transplantation ; ischemia-reperfusion ; tolerance ; rejection ; indoleamine-2,3-dioxygenase ; L-tryptophan ; amino acids ; MAC-T cell ; proteomics ; omics ; β-casein ; mTOR ; systemic inflammation ; dysbiosis ; gut ; microbiota ; obesity ; mice ; tyrosine ; cytokines ; behavior ; inflammation ; liver morphology ; color ; cell culture media ; LC-MS ; antioxidant ; cytotoxicity ; biomanufacturing ; 5-hydroxytryptamine ; secretion ; metabolism ; nitrofurantoin ; antibiotics ; human serum albumin ; molecular interactions ; FTIR ; fluorescence ; n/a ; bic Book Industry Communication::M Medicine ; bic Book Industry Communication::M Medicine::MM Other branches of medicine::MMG Pharmacology
    Language: English
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  • 9
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2023-03-07
    Description: The eradication of vector-borne diseases is threatened by the limited range of available insecticides, leading, inevitably, to the development of resistance. This is particularly concerning for malaria control, which relies heavily on insecticide-treated nets (ITNs) and indoor residual sprays (IRS). New chemistries are being developed, and innovative deployment of insecticides may play a role in overcoming resistance, either through new types of tools or new means of distribution. A variety of novel product types and vector control strategies are under development and evaluation, which is to be celebrated, but a strong evidence base is needed to guide effective operational deployment decisions. Novel approaches should be supported by robust data collected using appropriate and validated methods to monitor efficacy, durability, and any emerging resistance. This reprint presents original research into developing and characterizing new vector control products, as well as understanding and monitoring insecticide resistance. Review articles explore the impact of insecticide resistance and offer guidance on insecticide choice in the face of pyrethroid resistance. Consensus methodologies are presented, in the form of standard operating procedures (SOPs) designed to be adopted and used to generate reproducible data that can be compared and interpreted across and between studies. It is hoped that this collection of articles offers inspiration and guidance on how consistent data can be generated to inform more effective development, evaluation, and use of new and existing vector control tools.
    Keywords: prallethrin ; insecticide ; spatial treatment ; mosquito fitness ; protection ; pyrethroids ; Aedes albopictus ; Culex pipiens ; life tables ; mosquito ; bite-proof garment ; model ; textile ; non-insecticidal ; physical barrier ; insecticide selection ; out-crossing ; strain authentication ; laboratory screening ; pyrethroid ; pyrethroid resistance ; insecticide resistance ; insecticide resistance management ; vector control ; malaria ; malaria control ; Anopheles ; host-seeking behavior ; insecticide exposure ; pathogen transmission ; Aedes aegypti ; Anopheles gambiae ; ATSB ; Culex quinquefasciatus ; Iroquois ; RNAi ; Saccharomyces cerevisiae ; yeast ; Anopheles mosquito ; fertility ; ovary development ; pyriproxyfen (PPF) ; side-effects ; machine learning ; image classification ; automated identification ; convolutional neural network ; insecticide-treated net (ITN) ; PBO ITN ; synergist ITN ; dual-AI ITN ; insecticide resistance management (IRM) ; method validation ; durability monitoring ; bioinsecticide ; disease transmission ; insecticide-resistance ; mosquito-borne disease ; mosquito control ; natural compounds ; phytochemical ; malaria vector ; insecticide treated nets ; cytochrome P450s ; kdr ; cuticular resistance ; deltamethrin ; imidacloprid ; bifenthrin ; β-cyfluthrin ; etofenprox ; α-cypermethrin ; λ-cyhalothrin ; thiacloprid ; mosquitoes ; Attractive Toxic Sugar Bait (ATSB) ; Attractive Targeted Sugar Bait (ATSB) ; diagnostic bioassay ; resistance monitoring ; insecticide-treated nets (ITN) ; strain characterisation ; method development ; product evaluation ; quality control (QC) ; dual active ingredients (dual-AI) ; bioefficacy ; IRS ; application technology ; broflanilide ; clothianidin ; pirimiphos-methyl ; WHO tube ; WHO tunnel test ; ITNs ; interceptor ; interceptor G2 ; membrane ; human arm ; rabbit ; bioassay ; bio-efficacy ; n/a ; bic Book Industry Communication::G Reference, information & interdisciplinary subjects::GP Research & information: general ; bic Book Industry Communication::P Mathematics & science
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  • 10
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2023-05-11
    Description: Toxins are biologically active substances produced by most kinds of living organisms, bacteria, fungi, plants, and animals. They present a vast diversity of molecular structures and target a wide variety of receptors involved in a range of physiological processes. As toxins are selected during evolution to acquire/improve their disabling/lethal effects, they display finely tuned functional properties often associated with high affinities and selectivity. Moreover, toxins are valuable tools to unravel cellular processes due to their extreme specificity for cell surface and/or intracellular targets. Therefore, toxins are very attractive compounds because of their Janus-like character; while they mostly act as deadly poisons like monstrous Mr. Hyde, they can also be tamed into good remedies like admirable Dr. Jekyll. As such, they have been primarily investigated not only for the light they can throw on fundamental physiological processes but also for their potential therapeutic applications. This reprint, emerging from the 27th Annual Meeting of the French Society of Toxinology (SFET, http://sfet.asso.fr/international), will be of great interest for those in the scientific community who want to know more about the fascinating world of toxins.
    Keywords: toxins ; peptide chemistry ; native chemical ligation ; α-bungarotoxin ; click chemistry ; automated patch-clamp ; fluorescent peptide ; TE671 cells ; nicotinic acetylcholine receptor ; animal toxin ; bacterial toxin ; marine toxin ; medical application ; plant toxin ; toxin function/activity ; toxin receptor/target ; toxin structure ; Debaryomyces hansenii ; Wickerhamomyces anomalus ; Saccharomyces cerevisiae ; PDR transporters ; killer toxin ; fetal adrenomedullary chromaffin cell ; gambierol ; potassium currents ; calcium-activated K+ channels ; ATP-sensitive K+ channels ; catecholamine release ; Clostridium tetani ; Clostridium botulinum ; botulinum neurotoxin ; tetanus neurotoxin ; toxin gene regulation ; two-component system ; small RNA ; adenylate cyclase toxin ; Bordetella pertussis ; cyclic nucleotide ; cAMP ; spectrophotometric enzymatic assay ; ASIC ; sodium channels ; peptide ; PcTx1 ; APETx2 ; MitTx ; mambalgin ; pain ; nociception ; clostridial C3 toxin ; C3bot ; C3botE174Q ; dendritic cells ; macrophages ; monocytes ; stimulated emission depletion (STED) ; super-resolution microscopy ; trained immunity ; effector-triggered immunity ; effector-triggered trained immunity ; staphylococcal superantigen ; enterotoxin ; toxin pathogenicity ; immunomodulation ; molecular and cellular targets ; n/a ; bic Book Industry Communication::M Medicine ; bic Book Industry Communication::M Medicine::MM Other branches of medicine::MMG Pharmacology::MMGT Medical toxicology
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  • 11
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-04-11
    Description: Food waste is becoming an important and growing concern at both local and global levels. According to the Food and Agriculture Organization of the United Nations (FAO), one-third of all food production is wasted globally, and in particular, 1.3 billion tons of food produced for human consumption is wasted per year, representing an economic loss of EUR 800 billion. The main foods wasted are represented by vegetables, fruits, meat, and fish. Considering the high availability and the composition of food waste, there is an increasing interest in their bio-valorization. Moreover, according to the global Sustainable Development Goals (SDGs 12 and 13), an appropriate waste management represents an essential prerequisite for the sustainable development.This reprint collects interesting manuscripts regarding innovative research focused on food waste valorization through fermentation processes for obtaining value-added products such as enzymes, feed additives, biofuels, animal feeds as well as other useful chemicals or products, food-grade pigments, and single-cell protein (SCP), enhancing food security and environmentally sustainable development.
    Keywords: industrial food waste ; valorization ; biorefinery ; bioenergy ; biobased materials ; promotion policy ; rice husk ; pyrolysis ; porous biochar ; pore property ; surface composition ; microbial red pigment ; Monascus purpureus ; simultaneous hydrolysis and fermentation ; sustainability ; whey ; RSM ; bioethanol ; yeast fermentation ; sugar beet molasses ; industrial by-product ; scale-up ; agricultural waste ; wastewater ; microbial fuel cell ; techno-economic ; commercialization ; life cycle assessment ; Neurospora intermedia ; bread ; process development ; cheese whey ; Aspergillus awamori ; β-galactosidase ; lactose hydrolysis ; Acetobacter xylinum ; bacterial cellulose ; biosurfactant ; bioemulsifier ; waste frying oil ; Bacillus cereus ; food additives ; cookie ; microalgae ; DHA ; lignocellulosic biomass ; organosolv fractionation ; liquid fraction ; solid pulp ; omega-3 fatty acids ; soap ; olives ; olive oil ; fermentation ; food waste ; fish waste ; citrus peel ; aquafeed ; Saccharomyces cerevisiae ; Lactobacillus reuteri ; whey product ; proteins ; ultrafiltration ; nanofiltration ; keratinocytes scratch assay ; mozzarella cheese manufacturing ; pressing residue ; grape ; apple ; silage ; animal production ; enzyme production ; polyphenols ; Juglans regia L. ; walnut green husk ; agricultural wastes ; soil conditions ; glucans ; pectins ; Aspergillus oryzae ; rice hull ; paper mill wastewater ; bioremediation ; amylase ; solid-state fermentation (SSF) ; goat feeding ; durian peel ; silage additives ; propionate ; methane mitigation ; nitrogen balance ; waste management ; biofuel production ; circular economy ; single cell protein ; value-added product ; food and feed production ; yeast ; probiotics ; thema EDItEUR::T Technology, Engineering, Agriculture, Industrial processes::TB Technology: general issues ; thema EDItEUR::T Technology, Engineering, Agriculture, Industrial processes::TC Biochemical engineering::TCB Biotechnology
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  • 12
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-03-27
    Description: In this book, the performance of homogeneous and heterogeneous catalysts applied in biomass processing was assessed, paying special attention to the main advantages and challenges related to their use. Indeed, these challenges are opportunities to develop new research lines that could be fruitful in the near future. Thus, different studies are included, dealing with diverse subjects, with one main goal in common: the improvement of different aspects related to biomass processing through the use of catalysts.
    Keywords: nanospheroids ; zinc-doped CaO ; natural triglycerides ; aminolysis ; heterogeneous catalyst ; recyclability ; catalyst ; sodium hydroxide ; fatty acid methyl ester ; central composite rotatable design ; operational conditions ; aerated irrigation ; soil enzyme activity ; soil microbial biomass ; soil respiration ; bio-derived phenol ; Ni-Cu-Co/Al2O3 ; in-situ hydrodeoxygenation ; cyclohexane ; hydrogenolysis ; biomass ; 5-hydroxymethylfurfural ; 2,5-furandicrboxylic acid ; aerobic oxidation ; metal catalysts ; acid catalysis ; biodiesel ; biofuel ; esterification ; fatty acid ; methanolysis ; molybdenum oxide ; transesterification ; vegetable oil ; fatty acid methyl esters ; 2-ethyl-1-hexanol ; 1-heptanol ; 4-methyl-2-pentanol ; viscosity ; flash and combustion points ; methyl oleate ; methyl ricinoleate ; cellulase ; cellulose ; paper sludge ; Saccharomyces cerevisiae ; synergism ; furfural ; carbon-supported catalyst ; xylose conversion ; iron ; heterogeneous catalysts ; thermoset polymer ; epoxy ; cellulose nanofiber ; curing characteristics ; thermal properties ; mechanical properties ; RSM ; numerical optimization ; keratinase ; feather ; Bacillus sp. ; amino acids ; n/a ; thema EDItEUR::G Reference, Information and Interdisciplinary subjects::GP Research and information: general
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  • 13
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-03-27
    Description: Mitochondria play an increasingly central role in the context of cellular physiology. These organelles possess their own genome (mtDNA), which is functionally coordinated with the nuclear genome. Mitochondrial gene expression is mediated by molecular processes (replication, transcription, translation, and assembly of respiratory chain complexes) that all take place within the mitochondria. Several aspects of mtDNA expression have already been well characterized, but many more either are under debate or have yet to be discovered. Understanding the molecular processes occurring in mitochondria also has clinical relevance. Dysfunctions affecting these important metabolic ‘hubs’ are associated with a whole range of severe disorders, known as mitochondrial diseases. In recent years, significant progress has been made to understand the pathogenic mechanisms underlying mitochondrial dysfunction; however, to date, mitochondrial diseases are complex genetic disorders without any effective therapy. Current therapeutic strategies and clinical trials are aimed at mitigating clinical manifestations and slowing the disease progression to improve the quality of life of patients. The goal of the Special Issue ‘Mitochondria: from Physiology to Pathology’ published in Life (ISSN: 2075-1729) was to collect research and review articles covering the physiological and pathological aspects related to mtDNA maintenance and gene expression, mitochondrial biogenesis, protein import, organelle metabolism, and quality control.
    Keywords: atherosclerosis ; carotid intima-media thickness ; mitochondrial mutations ; cardiovascular risk factors ; mitochondria ; mtDNA ; cristae ; mitochondrial fission ; mitochondrial fusion ; mitochondrial diseas ; mitochondrial dynamics ; mitoenergetics ; mitosteroidogenesis ; LH ; cAMP ; Leydig cell ; mitochondrial DNA segregation ; heteroplasmy ; selective elimination ; mitophagy ; mitochondrial engineered nucleases ; kinases ; phosphorylation ; disease ; PINK1 ; Parkinson’s disease ; mitochondria homeostasis ; Cterm ; MELAS ; transmitochondrial cybrids ; aminoacyl-tRNA synthetases ; LARS2 ; mitochondrial disease ; therapeutic peptides ; FAD synthase ; FAD1 ; mitochondria localization ; Saccharomyces cerevisiae ; mRNA ; mitochondrial localization motif ; n/a ; thema EDItEUR::G Reference, Information and Interdisciplinary subjects::GP Research and information: general
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  • 14
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2022-02-01
    Description: Fermented foods are consumed all over the world and their consumption shows an increasing trend. They play many roles, from preservation to food security, improved nutrition and social well-being. Different microorganisms are involved in the fermentation process and the diversity of the microbiome is high.Fermented foods are food substrates that are invaded or overgrown by edible microorganisms whose enzymes hydrolyze polysaccharides, proteins and lipids to nontoxic products with flavors, aromas, and textures that are pleasant and attractive to the human consumer. Fermentation plays different roles in food processing, including the development of a wide diversity of flavors, aromas, and textures in food, lactic acid, alcoholic, acetic acid, alkaline and high salt fermentations for food preservation purposes, biological enrichment of food substrates with vitamins, protein, essential amino acids, and essential fatty acids and detoxification during food fermentation processing.
    Keywords: fermented foods ; nutritional guidelines ; legislation ; national food guides ; Saccharomyces cerevisiae ; biomass ; date extract ; optimization ; response surface methodology ; kinetic models ; antifungal ; bioprotection ; bread ; Lactobacillus plantarum ; phenyllactic acid ; Aspergillus ; Penicillium ; Fusarium ; sauerkraut ; microbiome ; fermentation ; probiotics ; high-throughput sequencing ; nutrition ; health benefits ; microbiology ; health ; bic Book Industry Communication::T Technology, engineering, agriculture::TB Technology: general issues
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  • 15
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-04-09
    Description: The purpose of this book was not to provide a comprehensive overview of the vast arena of how fungi and fungal metabolites are able to improve human and animal nutrition and health; rather, we, as Guest Editors, wished to encourage authors working in this field to publish their most recent work in this rapidly growing journal in order for the large readership to appreciate the full potential of wonderful and beneficial fungi. Thus, this Special Issue welcomed scientific contributions on applications of fungi and fungal metabolites, such as bioactive fatty acids, pigments, polysaccharides, alkaloids, terpenoids, etc., with great potential in human and animal nutrition and health.
    Keywords: fungal pigment ; natural dye ; spalting ; Scytalidium cuboideum ; dramada ; sustainable clothing ; selenium ; biofortification ; transporters ; mycorrhizal fungi ; plant growth-promoting rhizobacteria (PGPRs) ; fungal pigments ; textile dyeing ; toxicity testing ; biotechnological approaches ; challenges ; limits ; Saccharomyces boulardii ; Saccharomyces cerevisiae ; probiotics ; gastrointestinal tract ; Alginate ; β-glucan ; oligosaccharides ; elicitation ; Sargassum species ; Sparassis latifolia ; polyphenol ; antioxidant ; agave mezcalero bagasse ; apple bagasse ; solid-state fermentation ; secondary metabolites ; Pleurotus ostreatus ; Endophytic fungi ; Hyptis dilatata ; Pestalotiopsis mangiferae ; Pestalotiopsis microspora ; chemical elicitors ; antibacterial activity ; LC–ESI–Q–TOF–MS ; yeast ; biological control ; postharvest decay ; fruit ; mycorrhizae ; elevated CO2 ; Thymus vulgare ; growth ; photosynthesis ; metabolites ; biological activity ; Candida albicans ; non-albicans Candida species ; Candida auris ; aromatic alcohols ; fungi ; metabolomics ; NTCD ; additives ; functional foods ; nutraceuticals ; sustainability ; healthy aging ; Mortierella alpina ; animal fat by-product ; arachidonic acid ; ATR-FTIR spectroscopy ; Mucor circinelloides ; high-throughput screening ; metal ions ; phosphorus ; lipids ; biofuel ; FTIR spectroscopy ; bioremediation ; co-production ; natural colorants ; filamentous fungi ; stirred-tank bioreactor ; biodegradable films ; food package ; bioactive compounds ; FIP ; human health ; immunomodulation ; induced apoptosis ; lectin ; medicinal mushrooms ; polysaccharide ; terpenes and terpenoids ; melanin ; carotenoids ; polyketides ; azaphilones ; antitumor ; medical roles ; sphinganine-analog mycotoxins ; fumonisins ; AAL-toxin ; chemical structure ; toxicity ; genetics and evolution ; biosynthesis ; livestock ; ewes ; energy ; cytokines ; yeasts ; liquid swine diets ; MALDI-TOF ; biochemical identification ; growth temperature Ancom Gas Production System ; Candida krusei ; Candida lambica ; M. purpureus ; red yeast rice ; cholesterol reduction ; probiotic potential ; natural colorant ; extraction ability ; marine fungi ; Talaromyces albobiverticillius ; aqueous two-phases system extraction ; ionic liquids ; feed additive ; probiotic ; Sporidiobolus ruineniae ; tannase ; micro-fungi ; macro-fungi ; Ganoderma ; kombucha ; anticancer ; carotenoid ; medicinal mushroom ; mycobiome ; antimicrobial ; antifungal ; bioconversion ; cheese ; dairy ; Sclerotinia ; secondary metabolite ; endophytic fungi ; uncommon secondary metabolites ; thema EDItEUR::T Technology, Engineering, Agriculture, Industrial processes::TB Technology: general issues
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  • 16
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2022-02-24
    Description: The milk industry is largely based on dairy cattle production. After decades of great advancements in genetics, nutrition, and management, today, one cow can reach unprecedented levels of milk production. New challenges have been posed to preserving the health and welfare of these domestic animals. “High-Yielding Dairy Cows” is a collection of scientific papers focusing on three main areas: metabolic diseases, reproduction diseases, and herd (heath) management in confined and pasture production systems. This book aggregates knowledge from a molecular level to a more holistic approach on disease prevention and management, giving the reader an accurate overview of the current state of the art of this topic. It intends to contribute to ensuring the supply of ethical and responsible animal protein for about eight billion of people.
    Keywords: dairy cow ; fatty liver ; lipid metabolism ; oxidative stress ; SIRT1 ; dairy cows ; PPARγ ; non-alcoholic fatty liver disease (NAFLD) ; genetic factor ; dairy industry ; milking system ; work routine ; parlor ; milking model ; small dairy ; reproductive strategy ; parity ; season ; rank of AI ; type of AI ; heat stress ; whole transcript sequencing ; immune response ; stress response ; myostatin gene ; variation ; milk ; fatty acid ; cattle ; milk production ; metabolomics ; biomarkers ; flaxseed ; dry period ; enterolactone ; milk fatty acids ; peak of lactation ; lipolysis ; fatty acids ; casein ; postpartum diseases ; activin ; inhibin ; cytokines ; endometrium ; subclinical endometritis ; cow ; milk beta-hydroxybutyrate ; fat to protein content ratio ; left displaced abomasum ; negative energy balance ; alpha-tocopherol/vitamin E-related gene ; calving ; colostrum ; high-yield dairy cows ; inflammation ; health ; lactation ; liver ; mammary gland ; ultrasonography ; pregnancy proteins ; embryonic mortality ; fetal mortality ; body condition score ; urea ; β-hydroxybutyrate ; metabolism ; urea in milk ; primiparous cows ; lactation curves ; feeding system ; herd management ; protein metabolism ; amino acids ; milk protein ; Saccharomyces cerevisiae ; high-yield cows ; pH ; VFA ; inflammatory cytokines ; transition period ; ketosis ; RNA-Seq ; clustering ; liver metabolism ; Jersey ; oral calcium bolus ; calcium ; hypocalcemia ; mastitis ; culling ; reproduction ; herd health ; milking management ; production systems ; bic Book Industry Communication::M Medicine
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  • 17
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2022-10-25
    Description: Among gluten-related disorders, coeliac disease (CD) is the best-known one to date, a chronic immune-mediated enteropathy triggered by exposure to gluten in genetically predisposed individuals. It is a common disease, occurring at all ages and characterized by a wide spectrum of clinical manifestations, affecting any organ or tissue. The diagnosis rate of this pathology has increased in the last 10 years, so worldwide epidemiologic data are now available that show that CD is ubiquitous, with a prevalence of 1.4%, higher in female than male individuals. Currently, the only effective treatment for CD is strict and lifelong adherence to a gluten-free diet (GFD). However, CD research is changing rapidly due to the continuous advancing of knowledge. For this reason, the main goal of this Special Issue has been to address the existing knowledge gaps and help advance such important aspects as the pathophysiology, diagnosis, follow-up, and therapeutic options of this pathology. This Special Issue includes 12 peer-reviewed articles reporting on the latest research findings in and evidence related to CD. The published articles cover a range of topics central to CD and GFDs.
    Keywords: celiac disease ; relatives ; microbiota ; Saccharomyces cerevisiae ; Pseudomonas fluorescens ; Bacteroides caccae ; coeliac disease ; oral diseases ; oral prevention ; gingival bleeding ; sleep-related breathing disorders ; oral health ; enamel defects ; interceptive orthodontics ; data mining gluten free diet ; gluten proteins ; immunogenicity ; evidence-based practice ; case management ; treatment adherence and compliance ; anemia ; iron transporter ; IgA nephropathy ; tissue transglutaminase autoantibody ; tissue transglutaminase-targeted IgA deposits ; flow cytometry ; age ; sex ; lesion grade ; intraepithelial lymphocytes TCRγδ+ ; functional bowel disease ; gluten-free diet ; tissue biomarkers ; non-coeliac gluten sensitivity ; FODMAP diet ; dietitian ; rural health services ; gluten ; gliadin ; gluten immunogenic peptides ; non-dietary therapies ; gluten cross-contaminations ; dietary adherence ; vital gluten ; oat ; hidden gluten ; patients with CD ; symptoms ; gluten excretion urine ; gluten-free diet monitoring ; n/a ; bic Book Industry Communication::M Medicine
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  • 18
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2022-11-17
    Description: This Special Issue focuses on the effects of hydrostatic pressure on biological systems and the use of these effects for exploring the structure, function, and molecular dynamics of biological macromolecules and their ensembles. Here, we present a selection of papers highlighting new experimental findings and new theoretical concepts in high-pressure biosciences. In these studies, the authors combine pressure perturbation approaches with NMR and optical spectroscopy, kinetic and thermodynamic techniques, functional genomics and transcriptomics, and molecular dynamics simulations to gain new insights into the conformational dynamics of proteins and nucleic acids and to better understand the mechanisms of high-pressure adaptation in piezophiles. The articles collected in this issue demonstrate the unique exploratory potential of the pressure perturbation approach for biochemistry, biophysics, mechanistic enzymology, and evolutionary biology.
    Keywords: protein folding ; NMR ; high hydrostatic pressure ; thermodynamic stability ; protein–ligand binding ; high pressure ; Martian salts ; perchlorate ; BSA ; ANS ; viroid ; hydrostatic pressure ; temperature ; structure–activity relationship ; RNA World ; n/a ; G-quadruplex ; i-motif ; volumetric properties ; pressure-temperature phase diagram ; thermodynamics ; hepatitis B ; DNA ; oligo ; FRET ; FTIR ; spectroscopy ; pressure ; volume change ; TMPyP4 ; deep-sea adaptations ; compressibility ; cavities ; potential energy landscape ; yeast ; Saccharomyces cerevisiae ; high-pressure response ; genetic manipulation ; transcriptomics ; piezophysiology ; Anfinsen’s dogma ; native state N ; unfolded state U ; fibril state F ; protofibrils ; hen lysozyme ; circular dichroism ; 1H NMR spectroscopy ; atomic force microscopy ; cytochrome P450 reductase ; conformational change ; pressure-perturbation spectroscopy ; protein hydration ; reduction kinetics ; stop-flow spectroscopy ; Sorghum bicolor ; bic Book Industry Communication::G Reference, information & interdisciplinary subjects::GP Research & information: general ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences::PSB Biochemistry
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  • 19
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2022-02-01
    Description: Computational fluid dynamics (CFD), which uses numerical analysis to predict and model complex flow behaviors and transport processes, has become a mainstream tool in engineering process research and development. Complex chemical processes often involve coupling between dynamics at vastly different length and time scales, as well as coupling of different physical models. The multiscale and multiphysics nature of those problems calls for delicate modeling approaches. This book showcases recent contributions in this field, from the development of modeling methodology to its application in supporting the design, development, and optimization of engineering processes.
    Keywords: pumped hydroelectric storage ; inlet/outlet ; surrogate model selection ; multi-objective optimization process ; thermal environment ; numerical simulations ; ventilation cooling ; duct position ; the heat dissipation of LHD ; auxiliary ventilation ; triboelectric separation ; particle size distribution ; particle charge ; binary mixture ; in situ particle size measurement ; charge estimation ; computational fluid dynamics ; membrane module ; gas separation ; concentration polarization ; coal mining ; radon concentration ; ventilation ; occupational exposure assessment ; gasification ; fluidized bed ; CFD ; hydrodynamics ; multiphase flow ; surface tension modelling ; VOF ; rising bubbles ; capillary rise ; high pressure bubble column ; the critical bubble diameter ; the gas holdup ; the large bubbles ; the small bubbles ; Stirred fermenter ; dual-impeller ; Segment impeller ; Optimization ; rotating packed bed ; natural gas desulfurization ; droplet characteristic ; Eulerian–Lagrangian approach ; heat transport ; optimized design ; dynamic numerical simulation ; evaporative cooling system ; water recycling ; temperature ; humidity ; n/a ; gas–solid ; cyclone separator ; elevated temperature process ; pneumatic conveying ; large coal particles ; Euler–Lagrange approach ; DPM ; pressure drop ; swirling burner ; combustion characteristics ; industrial pulverized coal furnace ; scale-up ; scale-down ; Saccharomyces cerevisiae ; mechanistic kinetic model ; bioreactor ; concentration gradients ; digital twin ; bioprocess engineering ; bic Book Industry Communication::T Technology, engineering, agriculture::TB Technology: general issues
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  • 20
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2022-05-06
    Description: The Special issue "Biological and Pharmacological Activity of Plant Natural Compounds II" is continuing the intriguing research on the use of natural plant products. The second edition follows the aim of the first one.
    Keywords: Bergenia species ; botanical description ; traditional uses ; phytochemistry ; pharmacology ; anti-urolithiatic activity ; bergenin ; Flaxseed oil ; linusorb B3 ; anti-cancer ; apoptosis ; actin polymerization ; Src ; glioblastoma ; chlorogenic acid ; coffee ; cyclooxygenase ; espresso ; instant coffee ; platelet aggregation ; Rubia tinctorum L. ; antioxidants ; polyphenols ; ethylene glycol ; urolithiasis ; histophatology ; Saccharomyces cerevisiae ; β-glucan ; antimicrobial and anticancer activities ; detoxification ability ; immunomodulatory effect ; Aquilaria sinensis ; pheophorbide A ; MMP-2 ; MMP-9 ; HT-1080 ; advanced glycation end product (AGE) ; oxidative stress ; epithelial to mesenchymal transition ; AGE-inhibitor ; swertiamarin ; diabetic nephropathy ; astragaloside IV ; Astragalus membranaceus ; huang qi ; Astragali Radix ; liver ; liver regeneration ; 70% partial hepatectomy ; proliferation ; rat ; memory ; object recognition ; Ginkgo biloba ; dorsal hippocampus formation ; brain-derived neurotrophic factor ; Diclofenac ; γ-lactone ; nano-emulsion ; methylcellulose ; Ostrich oil ; Struthio camelus ; Caenorhabditis elegans ; leaf extract ; neuroprotection ; antioxidant activity ; DAF-16 ; Clerodendrum infortunatum ; terpenoids ; phenylpropanoids ; antidiabetic ; breast cancer ; Combretum indicum L. ; antidiabetic activity ; histopathology ; UPLC-QTOF/ESI-MS ; network pharmacology ; Biebersteinia heterostemon ; galegine ; hypotensive ; toxicity ; Sage ; Salvia officinalis ; cytotoxicity ; hepatoprotection ; MDA ; TAOxC ; MCF-7 ; HeLA cells ; HepG-2 cells ; Peganum harmala ; anti-inflammatory activity ; antioxidant ; LC-ESI-MS/MS ; traditional medicine ; rheumatoid arthritis ; rosmanol ; carnosol ; Callicarpa longissima ; TLR4/NF-κB/MAPK ; synergistic effect ; diabetes mellitus ; anti-diabetic drugs ; monoterpenes ; bic Book Industry Communication::M Medicine ; bic Book Industry Communication::M Medicine::MM Other branches of medicine::MMG Pharmacology
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  • 21
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2022-08-12
    Description: Mitochondria are the powerhouses of cells; however, mitochondrial dysfunction causes energy depletion and cell death in a variety of diseases. Altered oxidative phosphorylation and ion homeostasis are associated with ROS production resulting from the disassembly of respiratory supercomplexes and the disruption of electron transfer chains. In pathological conditions, the dysregulation of mitochondrial homeostasis promotes Ca2+ overload in the matrix and ROS accumulation, which induces the mitochondrial permeability transition pore formation responsible for mitochondrial morphological changes linked to membrane dynamics, and ultimately, cell death. Finally, studies on the impaired mitochondrial bioenergetics in pathology could provide molecular tools to counteract diseases associated with mitochondrial dysfunction.
    Keywords: aging heart ; Bcl-2 family ; mitochondria ; programmed cell death ; fatty acid oxidation ; palmitate ; oleate ; m.3243A&gt ; G mutation ; MT-ATP6 ; m.8909T&gt ; C ; ATP synthase ; nephropathy ; oxidative phosphorylation ; mitochondrial disease ; cardiolipin ; Barth syndrome ; Sengers syndrome ; respiratory chain ; Dilated Cardiomyopathy with Ataxia ; cardiomyopathy ; mammalian complex I ; NADH dehydrogenase ; complex I assembly ; complex I structure ; complex I deficiency ; supernumerary subunits ; electron transport chain ; mitochondrial dysfunction ; Leigh syndrome ; mitochondrial diseases ; yeast ; Saccharomyces cerevisiae ; pet mutants ; pancreatic endocrine cells ; mathematical model ; cellular bioenergetics ; diabetes ; glucagon ; insulin ; exercise ; immune system ; metabolic disease ; COVID-19 ; mitochondrial dynamics ; viral infections ; MAVS ; RIG-I ; MDA5 ; innate immune response ; SARS CoV-2 ; RSV ; influenza ; respiratory supercomplexes ; ROS ; ATP synthase/hydrolase ; mitochondrial permeability transition pore ; cristae ; cellular signaling ; human disease ; mitochondrial dynamic ; cell signaling ; cancer ; respiratory complexes ; oxidative stress ; mitochondrial DNA ; MTCYB mutations ; cytochrome b ; complex III ; aging ; energy metabolism ; entorhinal cortex ; lipoxidation-derived damage ; neurodegeneration ; oxidative damage ; protein import ; respiratory complex assembly ; supercomplexes ; mitochondrial proteostasis ; heart failure ; bioenergetics ; assembly factor ; atypical myopathy ; high-resolution respirometry ; toxicity assays ; cell culture ; equine primary myoblasts ; fibroblasts ; frozen tissue ; leukocytes ; oxygen consumption ; platelets ; respirometry ; skeletal muscle ; n/a ; bic Book Industry Communication::G Reference, information & interdisciplinary subjects::GP Research & information: general ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences ; bic Book Industry Communication::P Mathematics & science::PS Biology, life sciences::PSB Biochemistry
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  • 22
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2023-12-21
    Description: Almost 25 years ago, the first mammalian transient receptor potential (TRP) channel was cloned and published. TRP channels now represent an extended family of 28 members fulfilling multiple roles in the living organism. Identified functions include control of body temperature, transmitter release, mineral homeostasis, chemical sensing, and survival mechanisms in a challenging environment. The TRP channel superfamily covers six families: TRPC with C for “canonical”, TRPA with A for “ankyrin”, TRPM with M for “melastatin”, TRPML with ML for “mucolipidin”, TRPP with P for “polycystin”, and TRPV with V for “vanilloid”. Over the last few years, new findings on TRP channels have confirmed their exceptional function as cellular sensors and effectors. This Special Book features a collection of 8 reviews and 7 original articles published in “Cells” summarizing the current state-of-the-art on TRP channel research, with a main focus on TRP channel activation, their physiological and pathophysiological function, and their roles as pharmacological targets for future therapeutic options.
    Keywords: R5-920 ; n/a ; transient receptor potential channels ; photochromic ligands ; elementary immunology ; Purkinje cell ; EPSC ; substance P ; chemicals ; organ toxicity ; lymphocytes ; HSP70 ; physiology ; bioavailable ; inflammatory bowel disease ; platelets ; pollutants ; yeast ; regulatory T cells ; kinase ; Saccharomyces cerevisiae ; manganese ; cerebellum ; TRP channel ; NHERF ; inflammation ; nanoHPLC-ESI MS/MS ; TRPM7 ; chemical probes ; TRPM8 ; dorsal column nuclei ; TRPV2 ; TRPV3 ; calcitonin gene-related peptide ; TRPV1 ; ion channels ; transient receptor potential ; 2D gel electrophoresis ; MALDI-TOF MS(/MS) ; TRPV4 ; overproduction ; sulfur mustard ; oxidative stress ; graft versus host disease ; menthol ; topical ; chemosensor ; AP18 ; calcium signalling ; mucosal epithelium ; cuneate nucleus ; production platform ; TRPC channels ; ulcerative colitis ; channel structure ; xerostomia ; neutrophils ; cardiovascular system ; TRPC5 ; TRPC6 ; TRPC3 ; TRPC4 ; calcium signaling ; protein purification ; adipose tissue ; transient receptor potential (TRP) channels ; sodium ; TH17 ; diacylglycerol ; hypersensitivity ; TRPY1 ; GABAB ; HEK293 ; thrombosis ; ion channel ; TRPC ; pathophysiology ; SMAD ; toxicology ; endothelium ; calcium ; proteomics ; TRPA1 ; salivary glands ; TRP channels ; lipid mediators ; sensors ; radiation ; TRPM4 channel ; human medulla oblongata ; mGluR1 ; small molecules ; TRPC3 pharmacology ; bic Book Industry Communication::M Medicine
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  • 23
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-04-05
    Description: Yeasts are truly fascinating microorganisms. Due to their diverse and dynamic activities, they have been used for the production of many interesting products, such as beer, wine, bread, biofuels, and biopharmaceuticals. Saccharomyces cerevisiae (brewers’ or bakers’ yeast) is the yeast species that is surely the most exploited by humans. Saccharomyces is a top-choice organism for industrial applications, although its use for producing beer dates back to at least the 6th millennium BC. Bakers’ yeast has been a cornerstone of modern biotechnology, enabling the development of efficient production processes. Today, diverse yeast species are explored for industrial applications. This Special Issue “Yeast Biotechnology 2.0” is a continuation of the first Special Issue, “Yeast Biotechnology” (https://www.mdpi.com/books/pdfview/book/324). It compiles the current state-of-the-art of research and technology in the area of “yeast biotechnology” and highlights prominent current research directions in the fields of yeast synthetic biology and strain engineering, new developments in efficient biomolecule production, fermented beverages (beer, wine, and honey fermentation), and yeast nanobiotechnology.]
    Keywords: QH301-705.5 ; TP248.13-248.65 ; bioethanol production ; mead ; nanobiotechnology ; fermentation-derived products ; flavor ; citric acid production ; enzyme production ; non-Saccharomyces yeasts ; fermented beverages ; bioreactors ; Saccharomyces cerevisiae ; wine ; beer ; thema EDItEUR::P Mathematics and Science::PS Biology, life sciences
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  • 24
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-04-05
    Description: During the last few years, industrial fermentation technologies have advanced in order to improve the quality of the final product. Some examples of those modern technologies are the biotechnology developments of microbial materials, such as Saccharomyces and non-Saccharomyces yeasts or lactic bacteria from different genera. Other technologies are related to the use of additives and adjuvants, such as nutrients, enzymes, fining agents, or preservatives and their management, which directly influence the quality and reduce the risks in final fermentation products. Other technologies are based on the management of thermal treatments, filtrations, pressure applications, ultrasounds, UV, and so on, which have also led to improvements in fermentation quality in recent years. The aim of the issue is to study new technologies able to improve the quality parameters of fermentation products, such as aroma, color, turbidity, acidity, or any other parameters related to improving sensory perception by the consumers. Food safety parameters are also included.
    Keywords: QH301-705.5 ; Q1-390 ; TX341-641 ; low-ethanol wines ; wine-related fungi ; non-Saccharomyces ; yeasts ; narince ; wine quality ; tryptophol ; low ethanol wine ; serotonin ; non-conventional yeasts ; Bombino bianco ; Schizosaccharomyces pombe ; volatile compounds ; ethyl carbamate ; phthalates ; autochthonous ; meta-taxonomic analysis ; Pichia kluyveri ; pH control ; IAA ; Torulaspora delbrueckii ; chemical analyses ; aroma profile ; yeast ; enzymatic patterns ; wine flavor ; fermentation ; must replacement ; Saccharomyces cerevisiae ; malolactic fermentation ; wine ; HACCP ; food quality ; sequential inoculation ; alcoholic beverages ; itaconic acid ; biocontrol application ; white wine ; hydroxytyrosol ; tryptophan ; glucose ; kinetic analysis ; wine aroma ; amino acid decarboxylation ; lactic acid bacteria ; vineyard soil ; wine color ; tyrosol ; Saccharomyces ; Gompertz-model ; sequential culture ; biogenic amines ; SO2 reduction ; climate change ; Vineyard Microbiota ; A. terreus ; sulfur dioxide ; human health-promoting compounds ; Hanseniaspora guilliermondii ; non-Saccharomyces screening ; aromatic/sensorial profiles ; Malvar (Vitis vinifera L. cv.) ; probiotics ; Yeasts ; native yeast ; color ; glutathione ; hot pre-fermentative maceration ; technological characterization ; wine-related bacteria ; Riesling ; Torulaspora microellipsoides ; Lachancea thermotolerans ; Metschnikowia pulcherrima ; cashew apple juice ; resveratrol ; biocontrol ; shiraz ; Tannat ; ochratoxin A ; aroma compound ; trehalose ; wine composition ; Hanseniaspora uvarum yeast ; food safety ; acidity ; sensory evaluation ; viticulture ; melatonin ; alcoholic fermentation ; aroma ; thema EDItEUR::P Mathematics and Science::PS Biology, life sciences
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  • 25
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    MDPI - Multidisciplinary Digital Publishing Institute
    Publication Date: 2024-04-09
    Description: Yeasts are truly fascinating microorganisms. Due to their diverse and dynamic activities, they have been used for the production of many interesting products, such as beer, wine, bread, biofuels and biopharmaceuticals. Saccharomyces cerevisiae (bakers’ yeast) is the yeast species that is surely the most exploited by man. Saccharomyces is a top choice organism for industrial applications, although its use for producing beer dates back to at least the 6th millennium BC. Bakers’ yeast has been a cornerstone of modern biotechnology, enabling the development of efficient production processes for antibiotics, biopharmaceuticals, technical enzymes, and ethanol and biofuels. Today, diverse yeast species are explored for industrial applications, such as e.g. Saccharomyces species, Pichia pastoris and other Pichia species, Kluyveromyces marxianus, Hansenula polymorpha, Yarrowia lipolytica, Candida species, Phaffia rhodozyma, wild yeasts for beer brewing, etc. This Special Issue is focused on recent developments of yeast biotechnology with topics including recent techniques for characterizing yeast and their physiology (including omics and nanobiotechnology techniques), methods to adapt industrial strains (including metabolic, synthetic and evolutionary engineering) and the use of yeasts as microbial cell factories to produce biopharmaceuticals, enzymes, alcohols, organic acids, flavours and fine chemicals, and advances in yeast fermentation technology and industrial fermentation processes.
    Keywords: coffee processing ; coffee fermentation ; starter culture ; coffee beverage ; yeast ; Icewine ; Saccharomyces cerevisiae ; hyperosmotic stress ; CRISPR-Cas9 ; glycerol transport ; STL1 ; brewing ; Cyberlindnera ; NABLAB ; non-alcoholic beer ; non-conventional yeast ; non-Saccharomyces yeast ; response surface methodology ; Ustilago ; itaconic acid ; process improvement ; lignocellulosic feedstock ; yeasts ; grape ; federweisser ; wine ; microbiota identification ; MALDI-TOF MS Biotyper ; Torulaspora delbrueckii ; craft beer ; microbrewery plant ; mixed fermentation ; aroma profile ; strain collection ; aroma profiling ; gas chromatography ; wine yeast ; Saccharomyces ; fermentation ; volatile aroma compounds ; Simultaneous inoculation ; Alcoholic fermentation ; Malolactic fermentation ; Sacccharomyces cerevisiae ; Oenococcus oeni ; PN4TM ; OmegaTM ; Aroma profile ; antioxidant ; coffee ; W. anomalus ; industrial brewer’s strains ; adaptive laboratory evolution (ALE) ; snowflake phenotype ; beer fermentation ; wine yeasts ; lactic acid bacteria ; co-inoculation ; sequence inoculation ; flavor compounds ; color pigments ; cell printing ; piezoelectric dispensing ; GFP-tagged yeast clone collection ; living cell microarrays ; microfluidic chip ; dynamic single-cell analysis ; Candida albicans ; adhesion ; fibronectin ; nanomotion ; atomic force microscope (AFM) ; xylose metabolism ; genetic engineering ; biofuel ; Spathaspora passalidarum ; Pichia stipitis ; volatile organic compounds ; proton-transfer reaction-mass spectrometry ; Metschnikowia pulcherrima ; flavor ; non-Saccharomyces yeasts ; fermentation-derived products ; fermented beverages ; beer ; coffee bean fermentation ; itaconic acid production ; bioethanol production ; bioreactors ; yeast micro- and nanobiotechnology ; thema EDItEUR::T Technology, Engineering, Agriculture, Industrial processes::TB Technology: general issues
    Language: English
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  • 26
    ISSN: 1572-8773
    Keywords: major facilitator superfamily ; iron transport ; siderophores ; enterobactin ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract While in fungi iron transport via hydroxamate siderophores has been amply proven, iron transport via enterobactin is largely unknown. Enterobactin is a catecholate-type siderophore produced by several enterobacterial genera grown in severe iron deprivation. By using the KanMX disruption module in vector pUG6 in a fet3Δ background of Saccharomyces cerevisiae we were able to disrupt the gene YOL158c Sce of the major facilitator super family (MFS) which has been previously described as a gene encoding a membrane transporter of unknown function. Contrary to the parental strain, the disruptant was unable to utilize ferric enterobactin in growth promotion tests and in transport assays using 55Fe-enterobactin. All other siderophore transport properties remained unaffected. The results are evidence that in S. cerevisiae the YOL158c Sce gene of the major facilitator super family, now designated ENB1, encodes a transporter protein (Enb1p), which specifically recognizes and transports enterobactin.
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  • 27
    ISSN: 1432-0983
    Keywords: Key words Citrinin ; Pet mutants ; Mitochondrial biogenesis ; Vacuolar ATPase ; YKL118W disruption ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In countries with a hot climate the mycotoxin citrinin represents a serious problem in fungal food-poisoning. In humans the renal system is affected the most and the mitochondrial respiratory chain was identified as a possible sensitive target for this toxin. In addition, citrinin has an antifungal activity that also inhibits the growth of the yeast Saccharomyces cerevisiae. So far the precise mode of action and the subcellular targets for citrinin have not been identified. Therefore, we decided to use the model organism yeast for a genetic approach to identify genes that play a role in the sensitivity against this mycotoxin. A large collection of conditional respiratory deficient yeast mutants was screened for sensitivity against citrinin. One special pet-ts mutant was identified that exhibited a higher sensitivity against citrinin. The genetic system of yeast allowed the isolation of the respective wild-type gene. This yeast gene encodes the Vph2p subunit that is essential for the correct assembly of the vacuolar ATPase. Isolation of the mutated gene and gene-disruption experiments of VPH2 and the partially overlapping small YKL118W gene verified this finding. The wild-type VPH2 gene restores all defects of the mutants. In contrast to this, YKL118W gave no complementation and the null mutant showed no phenotype. Thereby the yeast vacuolar ATPase was found to be important for the toxic effect of citrinin in yeast cells. The consequences of this finding for the molecular mechanism of citrinin action and its relation to the mitochondrial respiratory chain are discussed.
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  • 28
    ISSN: 1432-0983
    Keywords: Key wordsPOL32 ; SRS2 ; DNA repair ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pol32 is a subunit of Saccharomyces cerevisiae DNA polymerase δ required in DNA replication and repair. To gain insight into the function of Pol32 and to determine in which repair pathway POL32 may be involved, we extended the analysis of the pol32Δ mutant with respect to UV and methylation sensitivity, UV-induced mutagenesis; and we performed an epistasis analysis of UV sensitivity by combining the pol32Δ with mutations in several genes for postreplication repair (RAD6 group), nucleotide excision repair (RAD3 group) and recombinational repair (RAD52 group). These studies showed that pol32Δ is deficient in UV-induced mutagenesis and place POL32 in the error-prone RAD6/REV3 pathway. We also found that the increase in the CAN1 spontaneous forward mutation of different rad mutators relies entirely or partially on a functional POL32 gene. Moreover, in a two-hybrid screen, we observed that Pol32 interacts with Srs2, a DNA helicase required for DNA replication and mutagenesis. Simultaneous deletion of POL32 and SRS2 dramatically decreases cellular viability at 15 °C and greatly increases cellular sensitivity to hydroxyurea at the permissive temperature. Based on these findings, we propose that POL32 defines a link between the DNA polymerase and helicase activities, and plays a role in the mutagenic bypass repair pathway.
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  • 29
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    Current genetics 38 (2000), S. 264-270 
    ISSN: 1432-0983
    Keywords: Key words Endopolygalacturonase ; Saccharomyces cerevisiae ; Kluyveromyces marxianus ; Pectinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene encoding endopolygalacturonase (EC 3.2.1.15) has been cloned, sequenced and expressed from three strains of Saccharomyces cerevisiae (including non-secretors) and three strains of Kluyveromyces marxianus. Both control and coding regions showed small differences within each species, one including loss of a potential glycosylation site. Two non-secreting S. cerevisiae strains (FY1679 and var. uvarum) had non-transcribed copies of functional genes. Maximum enzyme activity was achieved with the S. cerevisiae FY1679 gene in an expressing vector, with an enzyme activity of 51 μmol of reducing sugar released from polygalacturonic acid μg protein−1 min−1, the highest so far reported for a yeast.
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  • 30
    ISSN: 1432-0983
    Keywords: Key words Translation release factors ; Chromosome stability ; Microtubules ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chromosome stability in suppressor mutants for SUP35 and SUP45 genes coding for translation release factors was studied. We obtained spontaneous and UV-induced sup35 or sup45 mutants in a haploid strain disomic for chromosome III and tested the stability of an extra copy of this chromosome. The majority of the mutants showed increased chromosome instability. This phenotype was correlated with an increased sensitivity to the microtubule-poisoning drug benomyl which affects chromosome segregation at anaphase. Our data suggest that termination-translation factors eRF3 and eRF1 control chromosome transmission at mitotic anaphase in Saccharomyces cerevisiae.
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  • 31
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    Journal of thermal analysis and calorimetry 59 (2000), S. 643-648 
    ISSN: 1572-8943
    Keywords: drying ; intracellular water ; Saccharomyces cerevisiae ; TG
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The intracellular water content of a microorganism is an important parameter which is a determinant factor of its physiological properties. It is usually measured by complex and time consuming procedures. Thermogravimetry using infrared balance has been used for this purpose, through the identification of different drying steps occurring during the analysis. This work employs the same method with much smaller samples, using conventional thermogravimetric equipment in a simpler and faster way than other conventional procedures. Commercial yeast (Saccharomyces cerevisiae ) washed samples are analyzed in isothermal procedures which are run in about 30 min. The drying rate curve, when plotted as a function of the residual mass of the cells, allows the identification of the step where the intracellular water is lost and the determination of its content. The obtained values, on extracellular water free basis, are in the range of 65 to 69% and agree with those measured by other techniques.
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  • 32
    ISSN: 1573-4943
    Keywords: Homology modeling ; rotational energy barrier ; simulated annealing ; pyridoxal 5′-diphosphoadenosine ; pyridoxal 5′-triphosphoadenosine ; Saccharomyces cerevisiae ; phosphoenolpyruvate carboxykinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Molecular mechanics calculations have been employed to obtain models of the complexes between Saccharomyces cerevisiae phosphoenolpyruvate (PEP) kinase and the ATP analogs pyridoxal 5′-diphosphoadenosine (PLP-AMP) and pyridoxal 5′-triphosphoadenosine (PLP-ADP), using the crystalline coordinates of the ATP-pyruvate-Mn2+-Mg2+ complex of Escherichia coli PEP carboxykinase [Tari et al. (1997), Nature Struct. Biol. 4, 990–994]. In these models, the preferred conformation of the pyridoxyl moiety of PLP-ADP and PLP-AMP was established through rotational barrier and simulated annealing procedures. Distances from the carbonyl-C of each analog to ε-N of active-site lysyl residues were calculated for the most stable enzyme-analog complex conformation, and it was found that the closest ε-N is that from Lys290, thus predicting Schiff base formation between the corresponding carbonyl and amino groups. This prediction was experimentally verified through chemical modification of S. cerevisiae PEP carboxykinase with PLP-ADP and PLP-AMP. The results here described demonstrate the use of molecular modeling procedures when planning chemical modification of enzyme-active sites.
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  • 33
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    Molecular genetics and genomics 263 (2000), S. 81-89 
    ISSN: 1617-4623
    Keywords: Key words Flp recombinase ; Site-specific recombination ; Homologous recombination ; RAD52 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Site-specific recombination within the Saccharomyces cerevisiae 2-micron DNA plasmid is catalyzed by the Flp recombinase at specific Flp Recognition Target (FRT) sites, which lie near the center of two precise 599-bp Inverted Repeats (IRs). However, the role of IR DNA sequences other than the FRT itself for the function of the Flp reaction in vivo is not known. In the present work we report that recombination efficiency differs depending on whether the FRT or the entire IR serves as the substrate for Flp. We also provide evidence for the involvement of the IR in RAD52-dependent homologous recombination. In contrast, the catalysis of site-specific recombination between two FRTs does not require the function of RAD52. The efficiency of Flp site-specific recombination between two IRs cloned in the same orientation is about one hundred times higher than that obtained when only the two FRTs are present. Moreover, we demonstrate that a single IR can activate RAD52-dependent homologous recombination between two flanking DNA regions, providing new insights into the role of the IR as a substrate for recombination and a new experimental tool with which to study the molecular mechanism of homologous recombination.
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  • 34
    ISSN: 1617-4623
    Keywords: Key wordsYarrowia lipolytica ; Saccharomyces cerevisiae ; Ambient pH signalling ; Signal transduction ; Transmembrane protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Yarrowia lipolytica, the transcription factor Rim101p mediates both pH regulation and control of mating and sporulation. Like its homologues PacC of Aspergillus nidulans and Rim101p of Saccharomyces cerevisiae, YlRim101p is activated by proteolytic C-terminal processing, which occurs in response to a signal transduced by a pathway involving several PAL gene products. We report here the cloning and sequencing of two of these genes, PAL2 and PAL3. PAL2 encodes a putative 632-residue protein with six possible transmembrane segments, which differs from the transmembrane proteins Rim9p of S. cerevisiae and PalI of A. nidulans, but is homologous to A. nidulans PalH and to the product of the ORF YNL294c, a predicted polypeptide of unknown function in S. cerevisiae. PAL3 encodes an 881-residue polypeptide that is homologous to PalF of A. nidulans and to a newly identified putative polypeptide of S. cerevisiae. Both PAL2 and PAL3 are expressed constitutively, regardless of ambient pH. Mutations in these genes affect growth at alkaline pH and sporulation in both Y. lipolytica and in S. cerevisiae. They affect invasiveness of haploid strains in S. cerevisiae only, and conjugation in Y. lipolytica only. These results highlight the conservation of the Pal pathway initially described in A. nidulans.
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  • 35
    ISSN: 1617-4623
    Keywords: Key wordsGAL regulon ; Transcription ; Saccharomyces cerevisiae ; Galactose suppression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A plasmid clone that suppresses galactose toxicity in a gal7 yeast strain has been isolated from a multicopy genomic DNA library. Molecular analysis revealed that the region responsible for the suppression of galactose toxicity corresponds to the ORF YPR030w, which was named MRG19. A CEN-based plasmid carrying the above ORF was unable to suppress the toxicity. Galactokinase activity was substantially reduced in cell extracts obtained from transformants bearing multiple copies of MRG19. Multiple copies of MRG19 were also able to suppress galactokinase expression driven by the CYC1 promoter but not the TEF1 promoter. Multiple copies of MRG19 could not suppress GAL1-driven galactokinase expression in a gal80 strain. However, MRG19-mediated suppression of CYC1-driven galactokinase expression was independent of GAL80 function. These results imply that multiple copies of MRG19 suppress galactokinase expression probably at the level of transcription. In agreement with this idea, multiple copies of MRG19 also suppress β-galactosidase expression driven by the GAL1 promoter in a GAL80-dependent manner. Disruption of MRG19 leads to an increase in the cell density at stationary phase in synthetic complete medium. MRG19 encodes a previously uncharacterised 124-kDa protein that shows no sequence homology to any known proteins.
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  • 36
    ISSN: 1615-6110
    Keywords: Angiosperms ; Campanulaceae ; Wahlenbergia ; Breeding system ; pollination ; pollen collecting hairs ; autogamy ; self-compatibility ; nectar ; island biology ; Juan Fernández Islands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The reproductive biology ofW. berteroi, W. fernandeziana, and a putative hybrid betweenW. fernandeziana andW. grahamiae, endemic to Robinson Crusoe Island (Juan Fernández archipelago, Chile) was studied. Flowers are hermaphroditic, protandrous, offer nectar, and exhibit secondary pollen presentation involving pollen collecting hairs on the style. These features imply allogamy and biotic pollination. However, male and female phases overlap and no effective pollinators were observed. Experimental data indicate these taxa are self-compatible and facultatively autogamous, a conclusion also suggested by the pollen/ovule ratios. Selfing is accomplished when the stigmatic lobes reflex and touch the style, except forW. berteroi where they do not reflex completely. Autogamy is accomplished in the latter when pollen grains deposited on the inner surface of the corolla throat by the “pollen brush” are gathered by stigmatic lobes when shaken by wind. The degree of autogamy, and perhaps self-compatibility, seems to be inconstant, as implied by the variable natural seed set (overall range 21–188 seeds per fruit). A mixed mating system — primarily outcrossing/entomophilous, but also autogamous — must have been present in the continental ancestors of these taxa. Autogamy promoting self-fertilization is important now — on an island with scarce pollinators — and in the past — when the first founders arrived.
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  • 37
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    Plant systematics and evolution 222 (2000), S. 1-17 
    ISSN: 1615-6110
    Keywords: Angiosperms ; pollen ; pollen wall ; pollination ; exine ; intine ; aperture ; ornamentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Qualities of the stratified pollen walls were evaluated for their possible role in pollination (pollination modes, and pollen tube formation). The importance of studying pollen grains in their respective natural state is noted. Examples of pollen morphological features specific to pollination vectors are rare and difficult to demonstrate. However, some complex, but significant correlations are reported.
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  • 38
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    Plant systematics and evolution 222 (2000), S. 281-292 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Echium ; Esterhazya ; pollination ecology ; anthers ; pollen ; secondary pollen presentation ; sporopollenin ; viscin threads
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This paper reviews the origin, nature, systematic distribution, and the respective function of the highly variable and diverse thread-forming structures in angiosperm anthers (including somewhat similar, rare features in ferns and gymnosperms). On one hand, such threads may function as pollen-connecting vectors in forming pollen dispersal units, as sporopollenin threads (viscin threads), e.g. in Onagraceae, or sporopollenin-less threads in surprisingly many other angiosperm families. On the other hand, as is known from theImpatiens — “pollen basket”, threads or ropes may be involved in pollen presentation. In addition, for the first time two new examples of “pollen baskets” in Boraginaceae and Scrophulariaceae are reported. InEchium the basket is formed by cellular elements from the modified septal regions, whereas inEsterhazya a similar effect is achieved in an analogous manner by trichomes of the epidermal layer of the thecal wall. There is obviously a different function of these seemingly very similar baskets: inEchium the feature acts preferably as a pollen presentation agent, whereas inEsterhazya the primary function is to prevent all the pollen from being dispersed too soon.
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  • 39
    ISSN: 1617-4623
    Keywords: Key words Autonomously replicating sequence (ARS) ; Anti-bent DNA ; DNA structure ; Replication origin ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to better understand the involvement of the DNA molecule in the replication initiation process we have characterized the structure of the DNA at Autonomously Replicating Sequences (ARSs) in Saccharomyces cerevisiae. Using a new method for anti-bent DNA analysis, which allowed us to take into account the bending contribution of each successive base plate, we have investigated the higher-order structural organization of the DNA in the region which immediately surrounds the ARS consensus sequence (ACS). We have identified left- and right-handed anti-bent DNAs which flank this consensus sequence. The data show that this organization correlates with an active ACS. Analysis of the minimum nucleotide sequence providing ARS function to plasmids reveals an example where the critical nucleotides are restricted to the ACS and the right-handed anti-bent DNA domain, although most of the origins considered contained both left- and right-handed anti-bent DNAs. Moreover, mutational analysis shows that the right-handed form is necessary in order to sustain a specific DNA conformation which is correlated with the level of plasmid maintenance. A model for the role of these individual structural components of the yeast replication origin is presented. We discuss the possible role of the right-handed anti-bent DNA domain, in conjunction with the ACS, in the process of replication initiation, and potentialities offered by the combination of left- and right-handed structural components in origin function.
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  • 40
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    Molecular genetics and genomics 263 (2000), S. 877-888 
    ISSN: 1617-4623
    Keywords: Key words Staurosporine ; Vacuolar-type proton pumping ATPase ; Vacuolar protein sorting ; ATP-binding cassette transporter ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutations at several loci affect the sensitivity of the yeast Saccharomyces cerevisiae to staurosporine. We report here the characterization of novel staurosporine- and temperature-sensitive mutants (stt). Cloning and integration mapping showed that the genes STT2/STT6, STT5, STT7, STT8 and STT9 are allelic to VPS18, ERG10, GPI1, VPS34 and VPS11, respectively. The products of ERG10 and GPI1, respectively, catalyze mevalonate and glycosyl phosphatidylinositol anchor synthesis, while VPS18 and VPS11 genes belong to the class C VPS (Vacuolar Protein Sorting) genes, and the VPS34 gene is classified as a class D VPS. Therefore, staurosporine sensitivity is affected by ergosterol and glycolipid biosynthesis and by vacuolar functions. We found that other vps mutants belonging to classes C and D exhibit staurosporine sensitivity, and that they show calcium sensitivity and fail to grow on glycerol as the sole carbon source; both of the last two characteristics are shared by vacuolar H+-ATPase mutants (vma). As vma mutants were also found to show staurosporine-sensitive growth, staurosporine sensitivity is likely to be affected by acidification of the vacuole. Moreover, wild type yeast cells are more sensitive to staurosporine in alkaline media than in acidic media, suggesting that staurosporine is exported from the cytosol by H+/drug antiporters. Pleiotropic drug resistance (PDR) genes also provide some resistance to staurosporine, because Δpdr5, Δsnq2 and Δyor1 strains are more sensitive to staurosporine than the wild-type strain. This suggests that staurosporine is also exported by the ATP-binding cassette (ABC) transporters on the plasma membrane. vma mutants and vps mutants of classes C and D vps are sensitive to hygromycin B and vanadate, while ABC transporter-depleted mutants do not show such sensitivity, indicating that two systems differ in their ability to protect the cell against different types of drug.
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  • 41
    ISSN: 1617-4623
    Keywords: Key words DNA repair ; Helix-hairpin-Helix motif ; Methylmethane sulfonate (MMS) ; Saccharomyces cerevisiae ; UV radiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene MUS81 (Methyl methansulfonate, UV sensitive) was identified as clone 81 in a two-hybrid screen using the Saccharomyces cerevisiae Rad54 protein as a bait. It encodes a novel protein with a predicted molecular mass of 72,316 (632 amino acids) and contains two helix-hairpin-helix motifs, which are found in many proteins involved in DNA metabolism in bacteria, yeast, and mammals. Mus81p also shares homology with motifs found in the XPF endonuclease superfamily. Deletion of MUS81 caused a recessive methyl methansulfonate- and UV-sensitive phenotype. However, mus81Δ cells were not significantly more sensitive than wild-type to γ-radiation or double-strand breaks induced by HO endonuclease. Double mutant analysis suggests that Rad54p and Mus81p act in one pathway for the repair of, or tolerance to, UV-induced DNA damage. A complex containing Mus81p and Rad54p was identified in immunoprecipitation experiments. Deletion of MUS81 virtually eliminated sporulation in one strain background and reduced sporulation and spore viability in another. Potential homologs of Mus81p have been identified in Schizosaccharomyces pombe, Caenorhabditis elegans and Arabidopsis thaliana. We hypothesize that Mus81p plays a role in the recognition and/or processing of certain types of DNA damage (caused by UV and MMS) during repair or tolerance processes involving the recombinational repair pathway.
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  • 42
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    Journal of bioenergetics and biomembranes 32 (2000), S. 391-400 
    ISSN: 1573-6881
    Keywords: ATP synthase ; F1-ATPase ; Saccharomyces cerevisiae ; petite mutants ; epistasis ; mitochondrion ; pet mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The mitochondrial ATP synthase is a molecular motor that drives the phosphorylation ofADP to ATP. The yeast mitochondrial ATP synthase is composed of at least 19 differentpeptides, which comprise the F1 catalytic domain, the F0 proton pore, and two stalks, oneof which is thought to act as a stator to link and hold F1 to F0, and the other as a rotor.Genetic studies using yeast Saccharomyces cerevisiae have suggested the hypothesis thatthe yeast mitochondrial ATP synthase can be assembled in the absence of 1, and even 2, ofthe polypeptides that are thought to comprise the rotor. However, the enzyme complexassembled in the absence of the rotor is thought to be uncoupled, allowing protons to freelyflow through F0 into the mitochondrial matrix. Left uncontrolled, this is a lethal process andthe cell must eliminate this leak if it is to survive. In yeast, the cell is thought to lose ordelete its mitochondrial DNA (the petite mutation) thereby eliminating the genes encodingessential components of F0. Recent biochemical studies in yeast, and prior studies in E. coli,have provided support for the assembly of a partial ATP synthase in which the ATP synthaseis no longer coupled to proton translocation.
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  • 43
    ISSN: 1572-9699
    Keywords: electron microscopy ; killer effect ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A mesophilic wine yeast, Saccharomyces cerevisiae CSIR Y217 K − R − was subjected to the K2 killer effect of Saccharomyces cerevisiae T206 K + R + in a liquid grape medium. The lethal effect of the K2 mycoviral toxin was confirmed by methylene blue staining. Scanning electron microscopy of cells from challenge experiments revealed rippled cell surfaces, accompanied by cracks and pores, while those unaffected by the toxin, as in the control experiments, showed a smooth surface. Transmission electron microscopy revealed that the toxin damaged the cell wall structure and perturbed cytoplasmic membranes to a limited extent.
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  • 44
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    Antonie van Leeuwenhoek 78 (2000), S. 187-194 
    ISSN: 1572-9699
    Keywords: cAMP ; pseudohyphae ; Saccharomyces cerevisiae ; stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Saccharomyces cerevisiae pseudohyphae formation may be triggered by nitrogen deprivation and is stimulated by cAMP. It was observed that even in a medium with an adequate nitrogen supply, cAMP can induce pseudohyphal growth when S. cerevisiae uses ethanol as carbon source. This led us to investigate the effects of the carbon source and of a variety of stresses on yeast morphology. Pseudohyphae formation and invasive growth were observed in a rich medium (YP) with poor carbon sources such as lactate or ethanol. External cAMP was required for the morphogenetic transition in one genetic background, but was dispensable in strain Σ1278b which has been shown to have an overactive Ras2/cAMP pathway. Pseudohyphal growth and invasiveness also took place in YPD plates when the yeast was subjected to different stresses: a mild heat-stress (37 °C), an osmotic stress (1 m NACl), or addition of compounds which affect the lipid bilayer organization of the cell membrane (aliphatic alcohols at 2%) or alter the glucan structure of the cell wall (Congo red). We conclude that pseudohyphal growth is a physiological response not only to starvation but also to a stressful environment; it appears to require the coordinate action of a MAP kinase cascade and a cAMP-dependent pathway.
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  • 45
    ISSN: 1573-5028
    Keywords: gene expression ; heterologous expression ; H+/hexose symporter ; Lycopersicon esculentum ; quantitative PCR ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length (LeHT2) and two partial (LeHT1 and LeHT3) cDNA clones, encoding hexose transporters, were isolated from tomato (Lycopersicon esculentum) fruit and flower cDNA libraries. Southern blot analysis confirmed the presence of a gene family of hexose transporters in tomato consisting of at least three members. The full-length cDNA (LeHT2) encodes a protein of 523 amino acids, with a calculated molecular mass of 57.6 kDa. The predicted protein has 12 putative membrane-spanning domains and belongs to the Major Facilitator Superfamily of membrane carriers. The three clones encode polypeptides that are homologous to other plant monosaccharide transporters and contain conserved amino acid motifs characteristic of this superfamily. Expression of the three genes in different organs of tomato was investigated by quantitative PCR. LeHT1 and LeHT3 are expressed predominantly in sink tissues, with both genes showing highest expression in young fruit and root tips. LeHT2 is expressed at relatively high levels in source leaves and certain sink tissues such as flowers. LeHT2 was functionally expressed in a hexose transport-deficient mutant (RE700A) of Saccharomyces cerevisiae. LeHT2-dependent transport of glucose in RE700A exhibited properties consistent with the operation of an energy-coupled transporter and probably a H+/hexose symporter. The K m of the symporter for glucose is 45 μM.
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  • 46
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    Chemistry of natural compounds 36 (2000), S. 88-89 
    ISSN: 1573-8388
    Keywords: Saccharomyces cerevisiae ; yeast invertase ; active enzyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The substrate specificity of purified yeast invertase isolated fromSaccharomyces cerevisiae in transglycosylation reactions was determined. The enzyme is specific for primary alcohols. The yeast activity is a function of the alkyl length and substrate hydrophobicity (n-butyl, isobutyl, isoamyl alcohols).
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  • 47
    ISSN: 1572-8773
    Keywords: iron ; siderophores ; transport ; Saccharomyces cerevisiae ; fungi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Transport proteins of microorganisms may either belong to the ATP-binding cassette (ABC) superfamily or to the major facilitator (MFS)-superfamily. MFS transporters are single-polypeptide membrane transporters that transport small molecules via uniport, symport or antiport mechanisms in response to a chemiosmotic gradient. Although Saccharomyces cerevisiae is a non-siderophore producer, various bacterial and fungal siderophores can be utilized as an iron source. From yeast genome sequencing data six genes of the unknown major facilitator (UMF) family were known of which YEL065w Sce was recently identified as a transporter for the bacterial siderophore ferrioxamine B (Sit1p). The present investigation shows that another UMF gene, YHL047c Sce, encodes a transporter for the fungal siderophore triacetylfusarinine C. The gene YHL047c Sce (designated TAF1) was disrupted using the kanMX disruption module in a fet3 background (strain DEY 1394 Δfet3), possessing a defect in the high affinity ferrous iron transport. Growth promotion assays and transport experiments with 55Fe-labelled triacetylfusarinine C showed a complete loss of iron utilization and uptake in the disrupted strain, indicating that TAF1 is the gene for the fungal triacetylfusarinine transport in Saccharomyces cerevisiae and possibly in other siderophore producing fungi.
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  • 48
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    BioMetals 12 (1999), S. 289-294 
    ISSN: 1572-8773
    Keywords: accumulation ; gold ; proton efflux ; Saccharomyces cerevisiae ; toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract This paper examines the effects of ionic gold on Saccharomyces cerevisiae, as determined by long-term (growth in gold-containing media) and short-term interactions (H+ efflux activity). An increasing gold concentration inhibited growth and at 〈0.2 mM Au, growth was not observed. Transmission electron microscopy revealed no differences in ultrastructure but fine electron dense particles were observed in unstained preparations from gold-containing medium. After glucose addition (to 10mM) to starved suspensions of S. cerevisiae, glucose-dependent reduction of external pH occurred as the cells extruded protons. In the presence of increasing gold concentrations, the lag time before proton extrusion did not change but the rate and duration decreased significantly with a marked influence on proton efflux rate being observed at ≤ 10 μM. Extension of preincubation time of yeast cells in gold-containing medium resulted in a decreasing proton efflux rate and colloidal phase formation in the cell suspensions, the time between gold addition and the beginning of colloidal phase formation depending on the gold concentration used. Both Ca and Mg enhanced the inhibitory effect of gold on the yeast cells with Ca showing a stronger inhibitory effect than Mg.
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  • 49
    ISSN: 1432-0983
    Keywords: Key words Cysteine uptake ; Amino-acid permeases ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract Uptake by Saccharomyces cerevisiae of the sulphur-containing amino acid L-cysteine was found to be non-saturable under various conditions, and uptake kinetics suggested the existence of two or more transport systems in addition to the general amino-acid permease, Gap1p. Overexpression studies identified BAP2, BAP3, AGP1 and GNP1 as genes encoding transporters of cysteine. Uptake studies with disruption mutants confirmed this, and identified two additional genes for transporters of cysteine, TAT1 and TAT2, both very homologous to BAP2, BAP3, AGP1 and GNP1. While Gap1p and Agp1p appear to be the main cysteine transporters on the non-repressing nitrogen source proline, Bap2p, Bap3p, Tat1p, Tat2p, Agp1p and Gnp1p are all important for cysteine uptake on ammonium-based medium. Furthermore, whereas Bap2p, Bap3p, Tat1p and Tat2p seem most important under amino acid-rich conditions, Agp1p contributes significantly when only ammonium is present, and Gnp1p only contributes under the latter condition.
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  • 50
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    Current genetics 35 (1999), S. 77-81 
    ISSN: 1432-0983
    Keywords: Key words Adaptive mutations ; 6-N-hydroxylaminopurine ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract The frequency of reversion in a histidine-requiring mutant of Saccharomyces cerevisiae increases about ten-fold in stationary cells during histidine starvation. Histidine starvation enhances a similar frequency of reversion in a tryptophan-requiring mutant. Starvation, therefore, enhances mutation frequencies in a non-adaptive manner. The base analogue 6-N-hydroxylaminopurine (HAP) added prior to plating on medium with limited histidine strongly increases reversion of the histidine mutant. HAP-induced reversion increases further in stationary starving cells with the same kinetics as that which increases spontaneous reversion. Adding HAP to the stationary starving cells does not produce any effect.
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  • 51
    ISSN: 1432-0983
    Keywords: Key words Heteroduplex repair ; Strand discrimina-tion ; Strand interruptions ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract Site-directed mutagenesis was used to construct yeast centromere plasmids in which a strand nick or gap could be placed 5′ or 3′, on either strand, to a reporter gene (SUP4-o) carrying defined base mismatches. The plasmids were then transformed into yeast cells and the direction and efficiency of mismatch repair were assayed by scoring colouring of the transformant colonies. Strands that were nicked were consistently corrected more often than intact strands, but the effect was very small. However, placement of a small gap at the same positions as the nicks resulted in a marked increase in selection for the gapped strand and an enhanced efficiency of mismatch repair. Both the preference for the gapped strand and correction of the mismatch were offset by deletion of the mismatch repair gene PMS1. Together, the results suggest that strand interruptions can direct intracellular mismatch correction of plasmid-borne base mispairs in yeast.
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  • 52
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    Current genetics 36 (1999), S. 256-261 
    ISSN: 1432-0983
    Keywords: Key wordsFLO8 ; Transcriptional regulation ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract It is thought that the FLO8 gene encodes a transcriptional activator of the dominant flocculation gene FLO1 in Saccharomycescerevisiae. To determine other genes which are regulated by FLO8, a detailed comparison of the transcripts from the FLO8 and Δflo8 strains was carried out. In addition to the FLO1 gene, it was found that transcription of the FLO11 and STA1 genes is positively regulated by FLO8. In flo8 strains, not only transcripts of the FLO11, STA1, and FLO1 genes but also invasive growth, extracellular glucoamylase production, and flocculation were undetected. From these results, it is suggested that FLO8 regulates these characteristics via the transcriptional regulation of the FLO11, STA1, and FLO1 genes.
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  • 53
    ISSN: 1432-0983
    Keywords: Key words Psoralen sensitivity ; Cytochrome oxidase ; Saccharomyces cerevisiae ; Oxidative stress
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    Topics: Biology
    Notes: Abstract The yeast gene PSO7 was cloned from a genomic library by complementation of the pso7-1 mutant's sensitivity phenotype to 4-nitroquinoline-1-oxide (4NQO). Sequence analysis revealed that PSO7 is allelic to the 1.1-kb ORF of the yeast gene COX11 which is located on chromosome XVI and encodes a protein of 28-kDa localized in the inner mitochondrial membrane. Allelism of PSO7/COX11 was verified by non-complementation of 4NQO-sensitivity in diploids homo- and hetero-allelic for the pso7-1 and cox11::TRP1 mutant alleles. Sensitivity to 4NQO was the same in exponentially growing cells of the pso7-1 mutant and the cox11::TRP1 disruptant. Allelism of COX11 and PSO7 indicates that the pso7 mutant's sensitivity to photoactivated 3-carbethoxypsoralen and to 4NQO is not caused by defective DNA repair, but rather is due to an altered metabolism of the pro-mutagen 4NQO in the absence of cytochrome oxidase (Cox) in pso7-1/cox11::TRP1 mutants/disruptants. Lack of Cox might also lead to a higher reactivity of the active oxygen species produced by photoactivated 3-carbethoxypsoralen. The metabolic state of the cells is important for their sensitivity phenotype since the largest enhancement of sensitivity to 4NQO between wild-type (WT) and the pso7 mutant occurs in exponentially growing cells, while cells in stationary phase or growing cells in phosphate buffer have the same 4NQO resistance, irrespective of their WT/mutant status. Strains containing the pso7-1 or cox11::TRP1 mutant allele were also sensitive to the oxidative stress-generating agents H2O2 and paraquat. Mutant pso7-1, as well as disruptant cox11::TRP1, harboured mitochondria that in comparison to WT contained less than 5% and no detectable Cox activity, respectively.
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  • 54
    ISSN: 1432-072X
    Keywords: Key words Plasma membrane H+-ATPase ; PMA1 ; ATPase ; PMA2 ATPase ; Saccharomyces cerevisiae ; Copper stress ; Copper tolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The major yeast plasma membrane H+-ATPase is encoded by the essential PMA 1 gene. The PMA 2 gene encodes an H+-ATPase that is functionally interchangeable with the one encoded by PMA 1 , but it is expressed at a much lower level than the PMA 1 gene and it is not essential. Using genetically manipulated strains of Saccharomyces cerevisiae that exclusively synthesize PMA1 ATPase or PMA2 ATPase under control of the PMA1 promoter, we found that yeast cultivation under mild copper stress leads to a similar activation of PMA2 and PMA1 isoforms. At high inhibitory copper concentrations (close to the maximum that allowed growth), ATPase activity was reduced from maximal levels; this decrease in activity was less important for PMA2 ATPase than for PMA1 ATPase. The higher tolerance to high copper stress of the artificial strain synthesizing PMA2 ATPase exclusively, as compared to that synthesizing solely PMA1 ATPase, correlated both with the lower sensitivity of PMA2 ATPase to the deleterious effects of copper in vivo and with its higher apparent affinity for MgATP, and suggests that plasma membrane H+-ATPase activity plays a role in yeast tolerance to copper.
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  • 55
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    Molecular and cellular biochemistry 190 (1999), S. 47-54 
    ISSN: 1573-4919
    Keywords: calmodulin ; yeast calmodulin ; Ca2+ binding ; Ca2+ binding protein ; Saccharomyces cerevisiae ; interdomain interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Calmodulin of Saccharomyces cerevisiae has different Ca2+ binding properties from other calmodulins. We previously reported that the maximum number of Ca2+ binding was 3 mol/mol and the fourth binding site was defective, which was different from 4 mol/mol for others. Their macroscopic dissociation constants suggested the cooperative three Ca2+ bindings rather than a pair of cooperative two Ca2+ bindings of ordinary calmodulin. Here we present evidence for yeast calmodulin showing the intramolecular close interaction between the N-terminal half domain and the C-terminal half domain, while the two domains of ordinary calmodulin are independent of each other. We will discuss the relationship of the shape and the shape change caused by the Ca2+ binding to the enzyme activation in yeast. The functional feature of calmodulin in yeast will also be considered, which might be different from the one of vertebrate calmodulin.
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  • 56
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    Molecular and cellular biochemistry 202 (1999), S. 109-118 
    ISSN: 1573-4919
    Keywords: NF1 mutations ; IRA1 ; Saccharomyces cerevisiae ; RAS2 ; GAP activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The 2818 amino acids of neurofibromin, the product of the human NF1 gene, include a 230 amino acid Ras-GAP related domain (GRD). Functions which may be associated with the rest of the protein remain unknown. However, many NF1 mutations in neurofibromatosis 1 patients are found downstream of the GRD, suggesting that the C-terminal region of the protein is also functionally important. Since the C-terminal region of neurofibromin encompassing these mutations is homologous with the corresponding regions in the two Saccharomyces cerevisiae Ras-GAPs, Ira1p and Ira2p, we chose yeast as a model system for functional exploration of this region (Ira-C region). Three missense mutations that affect the Ira-C region of NF1 were used as a model for the mutagenesis of IRA1. The yeast phenotypes of heat shock sensitivity, iodine staining, sporulation efficiency, pseudohyphae formation, and GAP activity were scored. Even though none of the mutations directly affected the Ira1p-GRD, mutations at two of the three sites resulted in a decrease in the GAP activity present in ira1 cells. The third mutation appeared to disassociate the phenotypes of sporulation ability and GAP activity. This and other evidence suggest an effector function for Ira1p.
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  • 57
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    Molecular and cellular biochemistry 201 (1999), S. 17-24 
    ISSN: 1573-4919
    Keywords: Saccharomyces cerevisiae ; atomic force microscope ; bioscope ; organic synthesis ; molecular biology ; oxidative stress ; pore enlargement ; cell wall ; baker's yeast ; biotechnology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We imaged pores on the surface of the cell wall of three different industrial strains of Saccharomyces cerevisiae using atomic force microscopy. The pores could be enlarged using 10 mM diamide, an SH residue oxidant that attacks surface proteins. We found that two strains showed signs of oxidative damage via changes in density and diameter of the surface pores. We found that the German strain was resistant to diamide induced oxidative damage, even when the concentration of the oxidant was increased to 50 mM. The normal pore size found on the cell walls of American strains had diameters of about 200nm. Under conditions of oxidative stress the diameters changed to 400nm. This method may prove to be a useful rapid screening process (45-60 min) to determine which strains are oxidative resistant, as well as being able to screen for groups of yeast that are sensitive to oxidative stress. This rapid screening tool may have direct applications in molecular biology (transference of the genes to inside of living cells) and biotechnology (biotransformations reactions to produce chiral synthons in organic chemistry.
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  • 58
    ISSN: 1573-4943
    Keywords: Phosphoenolpyruvate carboxykinase ; oxaloacetate decarboxylase ; pyruvate kinase-like activity ; Anaerobiospirillum succiniciproducens ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Two members of the ATP-dependent class of phosphoenolpyruvate carboxykinases (PEPCKs) (Saccharomyces cerevisiae and Anaerobiospirillum succiniciproducens) have been comparatively studied with regard to their oxaloacetate (OAA) decarboxylase and pyruvate kinase-like activities. The pyruvate kinase-like activities were dependent on the presence of Mn2+; at the same concentrations Mg2+ was not effective. These activities were synergistically activated by a combination of both metal ions. V max for these activities in A. succiniciproducens and S. cerevisiae PEPCKs was 0.13% and 1.2% that of the principal reaction, respectively. The OAA decarboxylase activity was nucleotide independent and, with decreasing order of effectiveness, these activities were supported by Mn2+ and Mg2+. AMP is an activator of these reactions. V max for the OAA decarboxylase activities in A. succiniciproducens and S. cerevisiae PEPCKs was 4% and 0.2% that of the PEP-forming reaction, respectively.
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  • 59
    ISSN: 1617-4623
    Keywords: Key words Proteasome ; Synthetic lethality ; Saccharomyces cerevisiae ; AAA-ATPase ; 19S Regulatory particle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 19S regulatory particle of the yeast 26S proteasome consists of six related ATPases (Rpt proteins) and at least 11 non-ATPase proteins (Rpn proteins). RPN12 (formerly NIN1) encodes an Rpn component of the 19S regulatory particle and is essential for growth. To determine which subunit(s) of the 26S proteasome interact(s) with Rpn12, we attempted to screen for mutations that cause synthetic lethality in the presence of the rpn12-1 (formerly nin1-1) mutation. Among the candidates recovered was a new allele of RPT1 (formerly CIM5). This mutant allele was designated rpt1-2; on its own this mutation caused no phenotypic change, whereas the rpn12-1 rpt1-2 double mutant was lethal, suggesting a strong interaction between Rpn12 and Rpt1. The site of the rpt1-2 mutation was determined by DNA sequencing of the RPT1 locus retrieved from the mutant, and a single nucleotide alteration was found. This changes amino acid 446 of the RPT1 product from alanine to valine. The alanine residue is conserved in all Rpt proteins, except Rpt5, but no function has yet been assigned to the region that contains it. We propose that this region is necessary for Rpt1 to interact with Rpn12. The terminal phenotype of the rpn12-1 rpt1-2 double mutant was not cell cycle specific, suggesting that in the double mutant cells the function of the 26S proteasome is completely eliminated, thereby inducing multiple defects in cellular functions.
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  • 60
    ISSN: 1617-4623
    Keywords: Key wordsCAT8 ; Transcriptional regulation ; IDP2 ; JEN1 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract The yeast transcriptional activator Cat8p has been identified as a factor that is essential for the derepression of genes involved in gluconeogenesis (like FBP1, PCK1, ACR1, ICL1 and MLS1) when only non-fermentable carbon sources are provided. Cat8p-dependent expression is mediated by cis-acting elements in the respective promoters, which are named UAS/CSREs (upstream activating sequence/carbon source responsive element). To establish whether the function of Cat8p is restricted to the activation of gluconeogenesis or is also involved in the regulation of a greater variety of genes, we investigated the transcriptional regulation of two genes, IDP2 and JEN1, which exhibit a similar expression pattern to gluconeogenic genes, although IDP2 at least is not linked directly to the gluconeogenic pathway. We identified functional UAS/CSRE elements in the promoters of both genes. Expression studies revealed that JEN1 is regulated negatively by the repressors Mig1p and Mig2p, and that Cat8p is needed for full derepression of the gene under non-fermentative growth conditions. Furthermore, we showed that Mig2p is also involved in the repression of CAT8 itself. The results presented in this study support a model in which Cat8p-dependent gene activation is not restricted to gluconeogenesis, but targets a wide variety of genes which are strongly derepressed under non-fermentative growth conditions.
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  • 61
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    Molecular genetics and genomics 262 (1999), S. 589-599 
    ISSN: 1617-4623
    Keywords: Key words Ras/cAMP pathway ; Saccharomyces cerevisiae ; Snf1 ; Mig1 ; Mediator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyclin C and the cyclin C-dependent protein kinase are associated with the RNA polymerase II Mediator complex, which regulates initiation of transcription in response to signals from activators and repressors bound to upstream promoter elements. Disruption of the corresponding genes, SRB11 and SRB10, in budding yeast causes a reduction in expression of the GAL genes, which is particularly pronounced in a mig1 snf1 background. We have screened two yeast genomic libraries for genes that can suppress this phenotype when overexpressed. Seven suppressor genes were identified, GIS1–7. GIS1 encodes one of two related zinc-finger proteins, which also share two other highly conserved domains present in several eukaryotic transcription factors. GIS2 encodes a homologue of the mammalian CNBP and fission yeast Byr3 proteins. GIS3 and GIS4 predict proteins with no obvious similarities to any known proteins. GIS5–7 are identical to the previously described genes PDE2, SGE1 and TUB3, respectively. None of the suppressor genes seem to be involved in Mediator function. Instead, we find that the GIS1, GIS2 and GIS4 genes interact with the CDC25 gene, indicating a possible involvement of these genes in the RAS/cAMP signaling pathway.
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  • 62
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    Plant systematics and evolution 217 (1999), S. 43-53 
    ISSN: 1615-6110
    Keywords: Leguminosae ; Caesalpinia ; Angiosperms ; bee-pollination ; andromonoecy ; late-acting self-incompatibility ; fruiting success
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The pollination biology, breeding system and fruiting success ofCaesalpinia calycina andC. pluviosa var.sanfranciscana were studied in caatinga vegetation in Bahia, NE Brazil. The principal pollinators for both species were carpenter bees.Caesalpinia calycina is andromonoecious but inC. pluviosa all flowers are hermaphrodite. InC. calycina all selfed flowers were abscised within 72 h despite rapid self-pollen tube growth to the ovary and ovule penetration. Prevention of selfing therefore seems to be controlled by a post-zygotic mechanism. Both species had very low fruit-set and it is suggested that this is at least in part due to geitonogamous pollinations with ovule penetration by self pollen tubes.
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    Plant systematics and evolution 216 (1999), S. 265-288 
    ISSN: 1615-6110
    Keywords: Illiciaceae ; Illiciospermum ; Liriodendroidea ; Magnoliaceae ; Angiosperms ; Cretaceous ; fossil seeds ; Kazakhstan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cenomanian-Turonian sediments from the Sarbay locality in north-western Kazakhstan have yielded a rich assemblage of plant fossils including well preserved flowers, fruits, and seeds of angiosperms. This work describes fossil seeds assigned to theMagnoliaceae and theIlliciaceae. Three new species of the extinct magnoliaceous genusLiriodendroidea, L. asiatica, L. costata, andL. tenuitesta, are established and new information on the previously described species,L. alata, is provided. TheLiriodendroidea seeds are closely similar to seeds of extantLiriodendron, but are distinguished in being much smaller and winged. A new genus and species,Illiciospermum pusillum, is established based on seeds with close similarity to those of the extant genusIllicium. The seeds are small, anatropous and exotestal with outer epidermis of testa forming a palisade layer. The facets of the palisade cells have deeply undulate anticlinal walls. The micropyle area is seen on the outer integument as a transverse slit placed on a raised strophiole-like structure close to the hilum. TheIlliciospermum seeds represent the first unequivocal record of theIlliciaceae in the Cretaceous. Another seed of possible illiciaceous affinity is described as aff.Illiciospermum sp.
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  • 64
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    Plant systematics and evolution 214 (1999), S. 161-186 
    ISSN: 1615-6110
    Keywords: Rubiaceae ; Rubioideae ; Angiosperms ; cladistics ; DNA sequences ; phylogeny ; rps16 intron ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phylogeny of the subfamilyRubioideae (Rubiaceae) was estimated from sequence variation in therps16 intron (cpDNA) in 143 ingroup and 5 outgroup taxa. The analysis largely confirms a recent one based onrbcL sequences, but branch support is often much stronger. Three of the traditional subfamilies are supported,Rubioideae, Cinchonoideae s. str., andIxoroideae s. l. while there is no support forAntirheoideae. TheRubioideae are the sister group of all otherRubiaceae and comprise the tribesAnthospermeae, Coccocypseleae, Cruckshanksieae, Coussareeae, Gaertnereae, Hedyotideae, Knoxieae, Morindeae, Ophiorrhizeae, Paederieae, Pauridiantheae, Perameae, Psychotrieae, Rubieae, Spermacoceae, Theligoneae, andUrophylleae. TheHamelieae andHillieae belong to theCinchonoideae. Rachicallis andSiemensia should be transferred from theHedyotideae to theCinchonoideae. ThePauridiantheae, Urophylleae, Ophiorrhizeae, andRaritebe form the basalmost subclade of theRubioideae. The second basalmost clade consists of the generaLasianthus andPerama. The third basalmost clade consists of the tribesCoussareeae, Coccocypseleae andCruckshanksieae, and the generaDeclieuxia andHindsia. The tribesKnoxieae, Anthospermeae, Argostemmateae, Paederieae, Theligoneae, Rubieae, Hedyotideae, andSpermacoceae are members of one clade. TheKnoxieae are monophyletic ifOtiophora, Otomeria, andPentas are included. The tribeAnthospermeae is supported as monophyletic, but its subtribes are not. ThePaederieae, together withTheligonum, form a paraphyletic grade basal to theRubieae. TheHedyotideae, includingSchismatoclada, form a grade at the base of theSpermacoceae. TheGaertnereae are monophyletic and distinct from thePsychotrieae. TheMorindeae are monophyletic and includeDamnacanthus andMitchella. Schradera is the sister group of theMorindeae. ThePsychotrieae are monophyletic when theGaertnereae, Lasianthus, andDeclieuxia are excluded. The recognition of a subtribeHydnophytineae leaves the rest of thePsychotrieae paraphyletic.Psychotria is paraphyletic with respect to all other genera of the tribe. Approximately 50 genera are here classified for the first time based on molecular data.
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  • 65
    ISSN: 1615-6110
    Keywords: Campanulaceae ; Lobelia ; Angiosperms ; in situ hybridization ; karyotype evolution ; rDNA ; telomere
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three repeated DNA sequences (rDNA 5S, 18S-5.8S-26S and telomeric repeats) were localised in the genomes ofLobelia brasiliensis andL. imperialis var.kanitzii (subg.Tupa), both with 2n = 28, by fluorescence in situ hybridization (FISH). The results were used to analyse the genomic relationship between the species. With probe pTa71, the karyotypes of these species showed only one NOR site. Probe pTa794, which contains 5S rDNA, demonstrated differences between the species. Telomeric sequences, studied with probe pLT11, were not detected in ectopic sites, but different telomeres showed signals of varying intensity. Based on the results obtained, considerations are made on karyotype evolution inLobelia.
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  • 66
    ISSN: 1617-4623
    Keywords: Key words Cse1p ; Srp1p ; Importin ; Nuclear transport ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract The yeast Srp1p protein functions as an import receptor for proteins bearing basic nuclear localization signals. Cse1p, the yeast homolog of mammalian CAS, recycles Srp1p back to the cytoplasm after import substrates have been released into the nucleoplasm. In this report we describe genetic interactions between SRP1 and CSE1. Results from genetic suppression and synthetic lethality studies demonstrate that these gene products interact to ensure accurate chromosome segregation. We also describe new mutant alleles of CSE1 and analyze a new temperature-sensitive allele of CSE1, cse1-2. This allele causes high levels of chromosome missegregation and cell cycle arrest during mitosis at the nonpermissive temperature.
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    Molecular genetics and genomics 262 (1999), S. 332-341 
    ISSN: 1617-4623
    Keywords: Key words Leucine transport ; Saccharomyces cerevisiae ; Trifluoroleucine resistance ; LEP1 ; SAC3
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Leucine uptake by Saccharomyces cerevisiae is mediated by three transport systems, the general amino acid transport system (GAP), encoded by GAP1, and two group-specific systems (S1 and S2), which also transport isoleucine and valine. A new mutant defective in both group-specific transport activities was isolated by employing a gap1 leu4 strain and selecting for trifluoroleucine-resistant mutants which also showed greatly reduced ability to utilize l-leucine as sole nitrogen source and very low levels of [14C]l-leucine uptake. A multicopy plasmid containing a DNA fragment which complemented the leucine transport defect was isolated by selecting for transformants that grew normally on minimal medium containing leucine as nitrogen source and subsequently assaying [14C]l-leucine uptake. Transformation of one such mutant, lep1, restored sensitivity to trifluoroleucine. The complementing gene, designated LEP1, was subcloned and sequenced. The LEP1 ORF encodes a large protein that lacks characteristics of a transporter or permease (i.e., lacks hydrophobic domains necessary for membrane association). Instead, Lep1p is a very basic protein (pI of 9.2) that contains a putative bipartite signal sequence for targeting to the nucleus, suggesting that it might be a DNA-binding protein. A database search revealed that LEP1 encodes a polypeptide that is identical to Sac3p except for an N-terminal truncation. The original identification of SAC3 was based on the isolation of a mutant allele, sac3-1, that suppresses the temperature-sensitive growth defect of an actin mutant containing the allele act1-1. Sac3p has been previously shown to be localized in the nucleus. When a lep1 mutant was crossed with a sac3 deletion mutant, no complementation was observed, indicating that the two mutations are functionally allelic.
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  • 68
    ISSN: 1617-4623
    Keywords: Key wordsKluyveromyces lactis ; Saccharomyces cerevisiae ; GAL1 ; GAL80 ; Protein interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Gal1p carries out two functions in the galactose pathway of yeast. It activates Gal4p by interacting with Gal80p – a function that can also served by Gal3p – and it catalyzes the formation of galactose-1-phosphate. Recently, we and others have presented biochemical evidence for complex formation between Gal1p and Gal80p. Here, we extend these data and present genetic evidence for an interaction between Gal1p and Gal80p in vivo, using a two-hybrid assay. Interaction between Gal1p and Gal80p depends on the presence of galactose, but not on the catalytic activity of Gal1p. A new class of Kluyveromyces lactis mutants was isolated, designated Klgal1-m, which have lost the derepressing activity but retain galactokinase activity, indicating that the two Gal1p activities are functionally independent. The KlGal1-m proteins are defective in their ability to interact with Gal80p in a two-hybrid assay. The locations of gal1-m mutations identify putative interaction sites in Gal1p and Gal80p. A dominant mutation, KlGAL1-d, leads to a high level of constitutive expression of genes of the galactose pathway. The behavior of chimeric proteins consisting of Gal3p and KlGal1p sequences indicates that both the N-terminal and C-terminal halves of KlGal1p are involved in specific interaction with KlGal80p.
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  • 69
    ISSN: 1617-4623
    Keywords: Key words Oxidative stress signalling ; Mitochondria ; Pos9 (Skn7) ; Ccp1 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Saccharomyces cerevisiae two transcription factors, Pos9 (Skn7) and Yap1, are involved in the response to oxidative stress. Fusion of the Pos9 response-regulator domain to the Gal4 DNA-binding domain results in a transcription factor which renders the expression of a GAL1-lacZ reporter gene dependent on oxidative stress. To identify genes which are involved in the oxygen-dependent activation of the Gal4-Pos9 hybrid protein we screened for mutants that failed to induce the heterologous test system upon oxidative stress (fap mutants for factors activating Pos9). We isolated several respiration-deficient and some respiration-competent mutants by this means. We selected for further characterization only those mutants which also displayed an oxidative-stress-sensitive phenotype. One of the respiration-deficient mutants (complementation group fap6) could be complemented by the ISM1 gene, which encodes mitochondrial isoleucyl tRNA synthetase, suggesting that respiration competence was important for signalling of oxidative stress. In accordance with this notion a rho0 strain and a wild-type strain in which respiration had been blocked (by treatment with antimycin A or with cyanide) also failed to activate Gal4-Pos9 upon imposition of oxidative stress. Another mutant, fap24, which was respiration-competent, could be complemented by CCP1, which encodes the mitochondrial cytochrome c peroxidase. Mitochondrial cytochrome c peroxidase degrades reactive oxygen species within the mitochondria. This suggested a possible sensor function for the enzyme in the oxidative stress response. To test this we used the previously described point mutant ccp1 W191F , which is characterized by a 104-fold decrease in electron flux between cytochrome c and cytochrome c peroxidase. The Ccp1W191F mutant was still capable of activating the Pos9 transcriptional activation domain, suggesting that the signalling function of Ccp1 is independent of electron flux rates.
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  • 70
    ISSN: 1617-4623
    Keywords: Key words Gene expression ; Glycolysis ; GCR ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To determine whether similar regulatory mechanisms control the expression of glycolytic genes in yeast and human cells, we screened a human brain cDNA library for clones which complement the growth defect of the gcr2 mutant of Saccharomyces cerevisiae, and isolated hSGT1 (human suppressor of GCR two). Further work confirmed that the rescue of growth was associated with recovery of glycolytic enzyme activities, and that hSGT1 did not complement the growth defect of a gcr1 mutant. A hybrid protein comprising hSgt1p and the DNA-binding domain of Gal4p (GBD) activated a GAL1-lacZ reporter gene fusion, suggesting that the cloned gene may be a transcriptional activator. Two-hybrid experiments in yeast also indicate that hSgt1p interacts with Gcr1p. Northern analysis showed that hSGT1 is highly expressed in muscle and heart. Although the predicted amino acid sequence of hSgt1p does not display significant similarity to Gcr2p, we speculate that their functions may be analogous.
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  • 71
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    Molecular genetics and genomics 260 (1999), S. 551-558 
    ISSN: 1617-4623
    Keywords: Key wordsRAD54 ; Saccharomyces cerevisiae ; Recombination ; Mating-type ; DNA repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Homothallic Saccharomyces cerevisiae strains switch their mating-type in a specific gene conversion event induced by a DNA double strand break made by the HO endonuclease. The RAD52 group genes control recombinational repair of DNA double strand breaks, and we examined their role in native homothallic mating-type switching. Surprisingly, we found that the Rad54 protein was important but not essential for mating-type switching under natural conditions. As an upper limit, we estimate that 29% of the rad54 spore clones can successfully switch their mating-type. The RAD55 and RAD57 gene products were even less important, but their presence increased the efficiency of the process. In contrast, the RAD51 and RAD52 genes are essential for homothallic mating-type switching. We propose that mating-type switching in RAD54 mutants occurs stochastically with a low probability, possibly reflecting different states of chromosomal structure.
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  • 72
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; heterologous expression ; isoprenoids ; mevalonate diphosphate decarboxylase ; sterols ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sequence comparison with the mevalonate diphosphate decarboxylase (MVD) amino acid sequence of Saccharomyces cerevisiae identified an EST clone corresponding to a cDNA that may encode Arabidopsis thaliana MVD (AtMVD1). This enzyme catalyses the synthesis of isopentenyl diphosphate, the building block of sterol and isoprenoid biosynthesis, and uses mevalonate diphosphate as a substrate. Sequencing of the full-length cDNA was performed. The predicted amino acid sequence presents about 55% identity with the yeast, human and rat MVDs. The sequence of the genomic region of A. thaliana MVD was also obtained and Southern blot analysis on genomic DNA showed that A. thaliana could have at least one homologous MVD gene. In order to allow heterologous expression in S. cerevisiae, the MVD open reading frame (ORF) was then cloned under the control of the yeast PMA1 strong promoter. When expressed in yeast, the A. thaliana cDNA complemented both the thermosensitive MN19-34 strain deficient in MVD, and the lethal phenotype of an ERG19 deleted strain. However, the wild-type sterol content was not fully restored suggesting that the A. thaliana MVD activity may not be optimal in yeast. A two-hybrid assay was also performed to evaluate homodimer formation of the A. thaliana MVD and heterodimer formation between the plant and yeast heterologous enzymes.
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  • 73
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    Journal of bioenergetics and biomembranes 31 (1999), S. 95-104 
    ISSN: 1573-6881
    Keywords: F1-ATPase ; β-barrel domain ; mitochondria ; assembly ; yeast ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The crystal structure of mitochondrial F1-ATPase indicatesthat the α and β subunits fold into a structure defined by threedomains: the top β-barrel domain, the middle nucleotide-binding domain,and the C-terminal α-helix bundle domain (Abraham et al.1994); Bianchet et al., 1998). The β-barrel domains of theα and β subunits form a crown structure at the top ofF1, which was suggested to stabilize it (Abraham et al.1994). In this study. the role of the β-barrel domain in the α andβ subunits of the yeast Saccharomyces cerevisiae F1,with regard to its folding and assembly, was investigated. The β-barreldomains of yeast F1 α and β subunits were expressedindividually and together in Escherichia coli. When expressedseperately, the β-barrel domain of the β subunit formed a largeaggregate structure, while the domain of the α subunit waspredominately a monomer or dimer. However, coexpression of the β-barreldomain of α subunit domain. Furthermore, the two domains copurified incomplexes with the major portion of the complex found in a small molecularweight form. These results indicate that the β-barrel domain of theα and β subunits interact specifically with each other and thatthese interactions prevent the aggregation of the β-barrel domain of theβ subunit. These results mimic in vivo results and suggest thatthe interactions of the β-barrel domains may be critical during thefolding and assembly of F1.
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  • 74
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    World journal of microbiology and biotechnology 15 (1999), S. 629-630 
    ISSN: 1573-0972
    Keywords: Ethanol ; multi-drug resistance ; Saccharomyces cerevisiae ; trichothecin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Trichothecin-resistant mutants were isolated from saké yeast. These mutants were subjected to saké brewing, and showed a higher ethanol productivity than did the parents. They showed multidrug resistance, and resistance to organic compounds. We considered that the higher ethanol productivity of the mutants was related to their resistance to organic compounds and to their ethanol tolerance.
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  • 75
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    Plant molecular biology 39 (1999), S. 117-128 
    ISSN: 1573-5028
    Keywords: LEA protein ; osmotic stress ; Saccharomyces cerevisiae ; drought ; salt
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The biased amino acid composition and aperiodic (random coil) configuration of Group 1 late embryogenesis-abundant (LEA) proteins imply that these proteins are capable of binding large amounts of water. While Group 1 LEAs have been predicted to contribute to osmotic stress protection in both embryonic and vegetative tissues, biochemical support has been lacking. We have used Saccharomyces cerevisiae as a model system to test the putative osmoprotective function of a wheat Group 1 LEA protein, Em. We demonstrate that expression of Em protein in yeast cells is not deleterious to growth in media of normal osmolarity and attenuates the growth inhibition normally observed in media of high osmolarity. Enhanced growth is observed in the presence of a variety of osmotically active compounds indicating that Em protein is capable of mitigating the detrimental effect of low water potential in a relatively non-specific manner. These results are the first biochemical demonstration of an osmoprotective function for a Group 1 LEA and suggest that the yeast expression system will be useful in dissecting the mechanism of protection through structure-function studies.
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  • 76
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    World journal of microbiology and biotechnology 15 (1999), S. 561-564 
    ISSN: 1573-0972
    Keywords: α-Amylase ; fusion protein ; glucoamylase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A fusion gene containing the Bacillus subtilis α-amylase gene and Aspergillus awamori glucoamylase cDNA was expressed in Saccharomyces cerevisiae. The resulting bifunctional fusion protein having both α-amylase and glucoamylase activities secreted into the culture medium was purified to apparent homogeneity by affinity chromatography and gel filtration on Sephadex G-100. The enzyme had an apparent molecular mass of 150 kDa and showed an optimum pH and temperature of 6.0 and 60 °C, respectively. The main hydrolysis products from soluble starch were glucose and maltose.
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  • 77
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    Mycopathologia 142 (1998), S. 67-70 
    ISSN: 1573-0832
    Keywords: l-glutamine ; fructose-6-phosphate amidotransferase ; Candida albicans ; fungi ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; systemic mycoses chemotherapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The 3' part of the glucosamine-6-phosphate synthase gene from Histoplasma capsulatum was PCR amplified using degenerate primers designed from the known glucosamine-6-phosphate synthase gene sequences, cloned and sequenced. The computer analysis of the 676 bp sequence revealed the presence of two introns. The identities of the deduced amino acid sequence to the corresponding Saccharomyces cerevisiae and Candida albicans fragment are 65 and 63.8%, respectively.
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  • 78
    ISSN: 1572-9699
    Keywords: growth inhibition ; fatty acid composition ; Saccharomyces cerevisiae ; Yarrowia lipolytica ; Teucrium polium L. extract
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    Topics: Biology
    Notes: Abstract Aqueous Teucrium polium extract slightly inhibits the growth of Saccharomyces cerevisiae (Ki=0.029 [g/l]-1) and Yarrovia lipolytica (Ki=0.061 [g/l]-1). However, this extract causes important changes in the unsaturation degree (Δ/mol) of the cellular lipids. It moreover favours the increase of the linolenic acid concentration and the decrease of the oleic one in both species.
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  • 79
    ISSN: 1423-0127
    Keywords: Acquired immunodeficiency syndrome ; Human immunodeficiency virus ; Nef protein ; Myristylation ; Membrane permeabilisation ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The human immunodeficiency virus type 1 (HIV-1) Nef protein is essential for AIDS pathogenesis, but its function remains highly controversial. During stresses such as growth in the presence of copper or at elevated temperature, myristylated Nef is released from yeast cells and, after extended culture in stationary phase, it accumulates in the supernatant as a dense membranous material that can be centrifuged into a discrete layer above the cell pellet. This material is unique to Nef-producing cells and represents a convenient source of Nef that may have application in further biological studies. Within the yeast cell, electron microscopic examination shows that Nef localises in novel, membrane-bound bodies. These data support the evidence for a role of Nef in membrane perturbation and suggest that there may be a similar localisation for myristylated Nef in HIV-1 infected cells.
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  • 80
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    Current genetics 34 (1998), S. 269-279 
    ISSN: 1432-0983
    Keywords: Key words Double-strand breaks ; Heteroduplex DNA ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Spontaneous and double-strand break (DSB)-induced gene conversion in Saccharomyces cerevisiae was assayed using non-tandem chromosomal direct repeat crosses and plasmid × chromosome crosses. Each cross involved identical ura3 alleles marked with phenotypically silent restriction fragment length polymorphic (RFLP) mutations at approximately 100-bp intervals. DSBs introduced in vivo at HO sites in one allele stimulated recombination to Ura+ by more than two orders of magnitude. Spontaneous gene-conversion products were isolated from a related strain lacking a functional HO nuclease gene. The multiple markers did not appear to influence the frequency of direct repeat deletions for spontaneous or DSB-induced events. DSB-induced conversion reflected efficient mismatch repair of heteroduplex DNA. Conversion frequencies of equidistant markers on opposites sides of the DSB were similar in the direct repeat cross. In contrast, markers 5′ of the DSB (promoter-proximal) converted more often than 3′ markers in plasmid × chromosome crosses, a possible consequence of crossing-over associated with long conversion tracts. With direct repeats, bidirectional tracts (extending 5′ and 3′ of the DSB) occurred twice as often as in a plasmid × chromosome cross in which DSBs were introduced into the plasmid-borne allele. A key difference between the direct-repeat and plasmid×chromosome crosses is that the ends of a broken plasmid are linked, whereas the ends of a broken chromosome are unlinked. We tested whether linkage of ends influenced tract directionality using a second plasmid × chromosome cross in which DSBs were introduced into the chromosomal allele and found few bidirectional tracts. Thus, chromosome environment, but not linkage of ends, influences tract directionality. The similar tract spectra of the two plasmid × chromosome crosses suggest that similar mechanisms are involved whether recombination is initiated by DSBs in plasmid or chromosomal alleles.
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  • 81
    ISSN: 1432-0983
    Keywords: Key wordsPSO5/RAD16 ; Saccharomyces cerevisiae ; Nucleotide excision repair ; Oxidative stress ; Ribonucleotide reductase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of β-galactosidase from DNA damage-inducible RNR2-lacZ and RNR3-lacZ fusion constructs was compared in wild-type (WT) and pso5/rad16 mutant strains after treatment with five mutagens/oxidative stressors. While exposure to the mutagens UVC, 4NQO and H2O2 induced expression of the RNR2-lacZ and RNR3-lacZ fusion constructs in two WT strains, treatment with the two oxidative stressors tBOOH and paraquat did not. In the pso5-1 mutant induction of RNR2-lacZ was largely reduced after UVC and H2O2 while there was no significant induction of β-galactosidase expression after 4NQO treatment for this construct. For RNR3-lacZ there was strongly reduced expression of pso5-1 after UVC and 4NQO while H2O2 failed to induce expression of β-galactosidase. In the WT strains the ranking of the inducing power of the mutagens at 90% survival (as measured in the pso5-1 mutant) was 4NQO〉UVC〉H2O2. Though the WT strains were clearly more resistant that the pso5-1 mutant to the two oxidative stressors paraquat and tBOOH, these substances failed to significantly enhance expression of the RNR2-lacZ and RNR3-lacZ fusion constructs in both the WT and the pso5-1 mutant. Our data suggest that Pso5p/Rad16p has a function in the signal transducing pathway controlling DNA damage-inducible components of nucleotide excision repair.
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  • 82
    ISSN: 1432-0983
    Keywords: Key words Zinc-finger protein ; Nuclear localization ; Immuno electron microscopy ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In previous studies the AZF1 gene has been identified as a second high-copy number suppressor for a special mutant of the gene for the mitochondrial core enzyme of RNA polymerase. The first high-copy number suppressor of this mutant turned out to be the specificity factor MTF1 for mitochondrial transcription. Up to now, the influence of AZF1 on mitochondrial transcription, its precise localization in the cell and the regulation of its expression has not been determined. The putative protein contains a long stretch of poly-asparagine amino acids and a typical zinc-finger domain for DNA binding. These characteristic structural features were used to create the abbreviation AZF1 (Asparagine-rich Zinc Finger protein). An initial computer analysis of the sequence gave no conclusive results for the presence of a mitochondrial import sequence or a typical nuclear-targeting sequence. A recent more-detailed analysis identified a possible nuclear localization signal in the middle of the protein. Disruption of the gene shows no effect on plates with glucose-rich medium or glycerol. In this report a specific polyclonal antibody against Azf1p was prepared and used in cell-fractionation experiments and in electron-microscopic studies. Both of these clearly demonstrate that the AZF1 protein is localized exclusively in the nucleus of the yeast cell. Northern analysis for the expression of the AZF1 messenger RNA under different growth conditions was therefore performed to obtain new insights into the regulation of this gene. Together with the respective protein-expression analysis these data demonstrate that Azf1p is preferentially synthezised in higher amounts under non-fermentable growth conditions. Over-expression of Azf1p in the yeast cell does not influence the expression level of the mitochondrial transcription factor Mtf1p, indicating that the influence of Azf1p on the suppression of the special mitochondrial RNA polymerase mutant is an indirect one. Subcellular investigation of the deletion mutant by electron microscopy identifies specific ultrastructural cell-division defects in comparison to the wild-type.
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  • 83
    ISSN: 1432-0983
    Keywords: Key words Mitotic recombination ; DNA double-strand breaks ; Saccharomyces cerevisiae ; 8-Methoxypsoralen plus UVA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mitotic recombination within the ARG4 gene of Saccharomyces cerevisiae was analysed after treatment of cells with the recombinogenic agent 8-methoxypsoralen (8-MOP) plus UVA. The appearance of DNA double-strand breaks (DSBs) in the ARG4 region during post-treatment incubation was also tested. The results obtained after 8-MOP plus UVA treatment indicate that in mitotic cells: (1) recombination at the ARG4 locus is increased 30 – 500 fold per survivor depending on the strains and the doses employed, (2) the increase of recombination results essentially from gene conversion events which involve the RV site located in the 5′ region of the ARG4 gene twice as often as the Bgl site at the 3′ end, (3) depending on 8-MOP/UVA dose, ectopic gene conversion is associated with reciprocal translocation, (4) DSBs occur preferentially in the ARG 5′ region during post-treatment incubation, as well as in other intergenic regions containing both promoters or/and terminators of transcription, and (5) changes in sequence content in the 5′ region of ARG4, which influences positions and frequencies of DSBs formed during repair, are correlated with a modification of the local chromatin structure.
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  • 84
    ISSN: 1432-0983
    Keywords: Key wordsSaccharomyces bayanus ; Saccharomyces cerevisiae ; Translocation ; Speciation ; Duplicated gene ; RPL2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By a genomic comparison of two sibling yeasts, Saccharomyces bayanus and S. cerevisiae, we previously demonstrated that chromosomes II and IV of S. cerevisiae were rearranged into chromosomes 12 and 14 of S. bayanus or vice versa. In the present study we have delimited the translocation break sites in chromosomes II and IV by Southern hybridization using DNA fragments of S. cerevisiae cosmid clones as probes. The results suggest that the reciprocal translocation of chromosomes II and IV had occurred at duplicated RPL2 loci. Furthermore, the translocation sites in S. bayanus were confirmed by the cloning and sequence analysis of the regions flanking RPL2 loci. Several genes in the regions flanking the RPL2 loci were present in the order expected for a translocation at these loci between the two species. These results indicated that the reciprocal translocation between chromosomes II and IV was generated by homologous recombination at duplicated RPL2 loci on the two chromosomes. Therefore, we propose that duplicated genes or duplicated regions play an important role in altering genomic organization during the speciation of S. bayanus and S. cerevisiae.
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  • 85
    ISSN: 1432-0983
    Keywords: Key words Fructose-1 ; 6-bisphosphatase ; Catabolite repression ; Gluconeogenesis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have investigated the effect of different carbon sources and of different mutations on the capacity of two elements, UAS1 and UAS2, from the promoter of the FBP1 gene to form specific DNA-protein complexes and to activate expression of a reporter gene. The complexes are observed with nuclear extracts from yeast derepressed on glycerol or ethanol. When hxk2 mutants are grown on glucose the nuclear extracts are able to complex UAS1 but not UAS2, while for wild-type cells grown on galactose only the complex with UAS2 is formed. In contrast, in vivo the operation of both UASs is high in ethanol, moderate to low in glycerol, and negligible in galactose; no expression is observed in glucose even in a hxk2 background. There is no effect of a MIG1 deletion, either in the formation of DNA-protein complexes or on the expression of reporter genes.
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  • 86
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    Current genetics 34 (1998), S. 138-145 
    ISSN: 1432-0983
    Keywords: Key words Cytochrome c oxidase ; Saccharomyces cerevisiae ; Complex assembly
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report on the molecular and biochemical analysis of a set of 13 respiratory deficient mutants of Saccharomyces cerevisiae which are specifically altered in COX1, the gene encoding the subunit Cox1p of cytochrome c oxidase. DNA sequence analysis shows that three are due to frameshift mutations, two to nonsense mutations, and eight to missense mutations. All, except the missense mutant S157L, have impaired electron transfer and respiratory activity. Analysis of the mitochondrial translation products shows that when Cox1p is absent, Cox2p and Cox3p are still synthesized. In the missense mutants, the steady state levels in the mitochondrial membranes of the three mitochondrially encoded subunits Cox1p, Cox2p and Cox3p and the nuclear-encoded subunit Cox4p are reduced. In the frameshift and nonsense mutants, Cox1p is absent and Cox2p, Cox3p and Cox4p are considerably decreased or undetectable. A comparison of the steady state levels of Cox1p through Cox4p in the COX1, COX2, COX3 and COX4 mutants shows the interdependance of the accumulation of these four subunits in the mitochondrial membranes.
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  • 87
    ISSN: 1432-072X
    Keywords: Key words Plasma membrane H+-ATPase ; Saccharomyces cerevisiae ; Copper stress ; PMA1 ; PMA2 ; Gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of Saccharomyces cerevisiae exibited a more active plasma membrane H+-ATPase during growth in media supplemented with CuSO4 concentrations equal to or below 1 mM than did cells cultivated in the absence of copper stress. Maximal specific activities were found with 0.5 mM CuSO4. ATPase activity declined when cells were grown with higher concentrations up to 1.5 mM (the maximal concentration that allowed growth), probably due to severe disorganization of plasma membrane. Cu2+-induced maximal activation was reflected in an increase of V max (approximately threefold) and in the slight decrease of the K m for MgATP (from 0.93 ± 0.13 to 0.65 ± 0.16 mM). The expression of the gene encoding the essential plasma membrane ATPase (PMA1) was reduced with a dose-dependent pattern in cells grown with inhibitory concentrations of copper, while the weakly expressed PMA2 gene promoter was moderately more efficient in cells cultivated under mild copper stress (1.5-fold maximal activation). ATPase was activated by copper despite the slightly lower content of ATPase protein in the plasma membrane of Cu2+-grown cells and the powerful inhibitory effect of Cu2+ in vitro.
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    Molecular and cellular biochemistry 184 (1998), S. 67-79 
    ISSN: 1573-4919
    Keywords: Saccharomyces cerevisiae ; spheroplast ; permeabilization ; mitochondria ; oxidative phosphorylation ; porin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In this work, we first compared yeast mitochondrial oxidative metabolism at different levels of organization: whole cells (C), spheroplasts (S), permeabilized spheroplasts (PS) or isolated mitochondria (M). At present, S are more suitable for use than C for biochemical techniques such as fast extraction of metabolises and permeabilization. We show here that respiratory rates of S with various substrates are similar to C, which demonstrate that they are adapted to yeast bioenergetic studies. It appeared from ethanol metabolism ± NAD++ or NADH respiratory rates on PS that ethanol metabolism was largely cytosolic; moreover, the activity of NADH dehydrogenase was lesser in the case of PS than in S. By comparing PS and M, the biggest difference concerned the respiratory rates of pyruvate and pyruvate-malate, which were much lower for M. Thus mitochondria preparation caused an unidentified loss involved directly in pyruvate metabolism. When the respiratory rate was lowered as a consequence of a high kinetic control of oxidative activity upstream from the respiratory chain, a similar correlation between the increase in ATP/O and decrease in respiratory rate was observed. So, the intrinsic uncoupling of proton pumps is not a particularity of M. Secondly, we demonstrate the existence of a mechanism of retarded diffusion in yeast similar to that already observed in permeabilized mammalian cells for ADP. Such a mechanism also occurs in yeast for several respiratory substrates: the K0.5 for each substrate toward the respiration rate in PS always exceeds that for M. It is proposed that such a discrepancy is due to a restriction of metabolite movement across the outer mitochondrial membrane in permeabilized cells, i.e. regulation of the substrate permeability through porin channels. In the porin-deficient yeast mutant, the K0.5 for NADH is not significantly different in either M or PS and is comparable to that of the parent strain PS. This result confirms that this retarded diffusion is essentially due to the opening-closing of the porin channel.
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  • 89
    ISSN: 1573-4919
    Keywords: Saccharomyces cerevisiae ; NAD(P)H ; calcium ions ; cells immobilization ; oxygen consumption ; biotransformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The biochemical behaviour of four commercial strains of Saccharomyces cerevisiae was studied in the presence of calcium ions, acrylamide and bisacrylamide. Calcium ions at a concentration of 300 µM induced an increase of NAD(P)+ reduction in commercial Turkish and American strains, while in Chilean and Brazilian commercial strains, it diminished NAD(P)+ reduction. On the other hand, polyacrylamide monomers (acrylamide and bisacrylamide) induced a decrease of NAD(P)+ reduction in all strains studied in this paper. When membrane potential (ΔΨ) and oxygen consumption were measured in the presence of polyacrylamide monomers, a decrease of both was observed in all strains studied.
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  • 90
    ISSN: 1573-4919
    Keywords: Saccharomyces cerevisiae ; microorganisms ; dehydrogenases ; acetoacetate ; molecular modelling ; enantiomeric excess ; biotransformation ; baker's yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract This method gives a general ideal how to use crystallographic information of enzymes to understand reactions catalyzed by these biocatalysts, commonly used by biochemists to produce chiral products. The interactions of three acetoacetic esters with the enzymes L-lactate dehydrogenase and alcohol dehydrogenase were studied through molecular modelling computer program. These artificial substrates have been widely used to produce chiral synthons. Through this methodology it was possible to understand the conformational specificity of these enzymes with respect to the products and how these enzymes can be inhibited by modifying the structures of the artificial substrates. Also, it was possible to predict whether some type of artificial substrate will suffer reduction by cells that contain these dehydrogenases and what kind of configuration (R or S) the final product will have.
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  • 91
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    Plant cell reports 18 (1998), S. 143-147 
    ISSN: 1432-203X
    Keywords: Key wordsNicotiana tabacum ; Male germ unit ; Scanning electron microscopy ; Sperm isolation ; Angiosperms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sperm cells are released from pollen tubes of tobacco as linked cells, associated with the vegetative nucleus in an assemblage known as the male germ unit (MGU). Using light microscopy, the MGU assemblage appears to be ensheathed by cytoplasmic material of the pollen tube, which may stabilize their association. Following their release, the shape of the sperm cells and vegetative nucleus changes from an ellipsoidal to a more spheroidal morphology. When most of the cytoplasmic material is dispersed, a boundary remains around the two sperm cells. Using scanning electron microscopy, the cytoplasmic material surrounding the MGU appears filamentous, sometimes twisted and rope-like. Based on these observations, the function of the MGU of tobacco is discussed.
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  • 92
    ISSN: 1617-4623
    Keywords: Key words Mitochondrial protein sorting ; Processing of Cox2 ; Kluyveromyces lactis ; Leishmania major ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The small nuclear gene SOM1 of Saccharomyces cerevisiae was isolated as a multicopy suppressor of a mutation in the IMP1 gene, which encodes the mitochondrial inner membrane peptidase subunit 1 (Imp1). Analysis revealed that Som1 and Imp1 are components of a mitochondrial protein export system, and interaction between these two proteins is indicated by the genetic suppression data. Here we describe the identification of a gene from Kluyveromyces lactis, which restores respiratory function to a S. cerevisiae SOM1 deletion mutant at 28° C. The sequence of the K. lactis gene predicts a protein product of 8.1-kDa, comprising 71 amino acid residues, with a putative mitochondrial signal sequence at its N-terminus. The protein is 50% identical to its S.cerevisiae counterpart. The expression pattern of a homologous sequence in Leishmania major suggests a more general role for SOM1 in mitochondrial biogenesis and protein sorting. The various Som1 proteins exhibit a highly conserved region and a remarkable pattern of cysteine residues. A protein of the expected size was transcribed and translated in vitro. The Som1 protein was detected in fractions of S. cerevisiae enriched for mitochondria and found to be associated with the inner mitochondrial membrane.
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  • 93
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    Molecular genetics and genomics 260 (1998), S. 417-425 
    ISSN: 1617-4623
    Keywords: Key words Centromere and promoter factor 1 (Cpf1p) ; Protein-protein interaction ; Saccharomyces cerevisiae ; Environmental adaptations ; Transcriptional activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcriptional regulation of the yeast cytochrome c 1 gene (CYT1) in response to oxygen and carbon source is mediated by Hap1p and the Hap2 complex. Furthermore, the centromere-binding factor 1 (Cbf1p) associates with the CYT1 upstream region (UASCYT1), but its direct activation potential is insignificant. The possible role of Cbf1p as a modulator of transcriptional adaptation to changes in nutritional conditions was examined. In electrophoretic mobility shift assays (EMSA) using yeast nuclear extracts, Cbf1p was found to exist as homo- and heterodimers of processed subforms of 54 and 37 kDa. An additional 18-kDa version was the only species found in anaerobic cells grown under an atmosphere of purified nitrogen, but not when CO2 was used to establish anaerobiosis. All three dimers of the 37 and 54 kDa versions of Cbf1p that occurred in oxidatively growing cells gave rise to hetero-oligomeric complexes containing other as yet unidentified protein(s). Complex formation was not observed with extracts from cultures grown on high levels of glucose and was dependent on pre-assembly in the absence of target DNA. Pre-treatment with alkaline phosphatase enhanced formation of these higher-order complexes. The C-terminal 18-kDa segment of Cbf1p, which can undergo dimerization and bind DNA, does not induce supershifts after preincubation and is not influenced by dephosphorylation. We propose that the N-terminal domain is subject to carbon source- or growth-dependent phosphorylation/dephosphorylation events that result in differential recruitment of additional factors to promoters of genes that encode proteins required for non-fermentative growth.
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  • 94
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    Molecular genetics and genomics 260 (1998), S. 102-107 
    ISSN: 1617-4623
    Keywords: Key words Immunosuppressant ; Uracil permease ; FUR4 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The immunosuppressant leflunomide inhibits cytokine-stimulated proliferation of lymphoid cells in vitro and also inhibits the growth of the eukaryotic microorganism Saccharomyces cerevisiae. To elucidate the molecular mechanism of action of the drug, two yeast genes which suppress the anti-proliferative effect when present in multiple copies were cloned and designated MLF1 and MLF2 for multicopy suppressor of leflunomide sensitivity. DNA sequencing analysis revealed that the MLF1 gene is identical to the FUR4 gene, which encodes a uracil permease and functions to import uracil efficiently. The MLF2 was found to be identical to the URA3 gene. Excess exogenous uracil also overcomes the anti-proliferative effect of leflunomide on yeast cells. Uracil prototrophy also conferred resistance to leflunomide. Uracil uptake was inhibited by leflunomide. Thus, the growth inhibition by leflunomide seen in a S. cerevisiae ura3 auxotroph is due to the inhibition of the entry of exogenous uracil via the Fur4 uracil permease.
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  • 95
    ISSN: 1617-4623
    Keywords: Key words Cell cycle ; mRNA splicing ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The S. cerevisiae CDC40 gene was originally identified as a cell-division-specific gene that is essential only at elevated temperatures. Cells carrying mutations in this gene arrest with a large bud and a single nucleus with duplicated DNA content. Cdc40p is also required for spindle establishment or maintenance. Sequence analysis reveals that CDC40 is identical to PRP17, a gene involved in pre-mRNA splicing. In this paper, we show that Cdc40p is required at all temperatures for efficient entry into S-phase and that cell cycle arrest associated with cdc40 mutations is independent of all the known checkpoint mechanisms. Using immunofluorescence, we show that Cdc40p is localized to the nuclear membrane, weakly associated with the nuclear pore. Our results point to a link between cell cycle progression, pre-mRNA splicing, and mRNA export.
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  • 96
    ISSN: 1617-4623
    Keywords: Key words Protein kinase C ; Signal transduction ; Transposon mutagenesis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We employed the constitutive BCK1-20 allele of the gene for the MAP kinase kinase kinase (MAPKKK) in the yeast Pkc signal transduction pathway to develop a genetic screen for mutants in genes encoding upstream components. Transposon mutagenesis yielded a mutant that was completely dependent on the active allele in the absence of osmotic stabilization. The transposon had integrated at the yeast SLG1 (HCS77) locus. This gene encodes a putative membrane protein. Haploid slg1 deletion strains are sensitive to caffeine, as expected for mutants in the Pkc pathway, as well as a variety of other drugs. The response to elevated temperatures and the dependence on osmotic stabilization depends on the genetic background. Thus, in the strain used for mutagenesis, disruption of SLG1 causes the cells to become non-viable in the absence of osmotic stabilization at both 30° C and 37° C. In a different genetic background this phenotype was not observed. Sensitivity of the haploid deletion mutants to caffeine can be partially suppressed by overexpression of genes for other components of the Pkc pathway, such as PKC1, SLT2, ROM2, and STE20. In addition, a SLG1-lacZ reporter construct shows higher expression in the presence of caffeine or magnesium chloride in a wild-type diploid background.
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  • 97
    ISSN: 1617-4623
    Keywords: Key words Dual-specificity phosphatase ; DNA synthesis ; Telophase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Cdc14 protein encodes a dual-specificity protein phosphatase which functions in late mitosis, and considerable genetic evidence suggests a role in DNA replication. We find that cdc14 mutants arrested in late mitosis maintain persistent levels of mitotic kinase activity, suggesting that Cdc14 controls inactivation of this kinase. Overexpression of Sic1, a cyclin-dependent protein kinase inhibitor, is able to suppress telophase mutants such as dbf2, cdc5 and cdc15, but not cdc14. It does, however, force cdc14-arrested cells into the next cell cycle, in which an apparently normal S phase occurs as judged by FACS and pulsed-field gel electrophoretic analysis. Furthermore, in a promoter shut-off experiment, cells lacking Cdc14 appear to carry out a normal S phase. Thus Cdc14 functions mainly in late mitosis and it has no essential role in S phase.
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  • 98
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    Molecular genetics and genomics 258 (1998), S. 512-520 
    ISSN: 1617-4623
    Keywords: Key words Homologous recombination ; Double-strand breaks ; Recombination intermediate ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In most eukaryotic organisms, recombination events leading to exchanges between homologous chromosomes link the homologs in a manner that allows their proper attachment to the meiotic spindle. In the yeast Saccharomyces cerevisiae these exchanges are initiated in early prophase as double-strand breaks in the DNA. These breaks are processed through a series of intermediates to yield mature crossovers late in prophase. The following experiments were designed to monitor the appearance of the earliest recombinant DNA strands formed in this process. A polymerase chain reaction assay was devised that allows the detection of recombinant strands at a known initiation site for meiotic recombination. The time and rate of appearance of recombinant strands was found to coincide with commitment to recombination, demonstrating that DNA strands bearing sequences from both parental chromosomes are rapidly formed after the initiation of meiotic recombination.
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  • 99
    ISSN: 1615-6102
    Keywords: GeneTUB2 ; Saccharomyces cerevisiae ; Antibody TU-14 ; Cortical β-tubulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The distribution of microtubules inSaccharomyces cerevisiae was studied with the monoclonal antibody (mab) TU-14 against β-tubulin. Immunoblotting and immunoprecipitation experiments with a strain overexpressing Tub2p confirmed that the mab TU-14 specifically recognized Tub2p. By immunofluorescence microscopy, the mab TU-14 attached to all known tubulin structures labelled with the standard polyclonal anti-β-tubulin antibody 206-1. In addition, the mab TU-14 revealed cortical patches in wild-type cells and an abundant network of fibres in the cortex of spheroplasts cultivated in nutrient medium. These cortical fibres seemed to be specific to spheroplasts and suggest that the accumulated Tub2p is predominantly associated with the plasma membrane.
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  • 100
    ISSN: 1573-4986
    Keywords: Saccharomyces cerevisiae ; oligosaccharide structure ; antigenic glycoprotein ; mannan ; allergens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Mannans of the yeast Saccharomyces cerevisiae have been implicated as containing the allergens to which bakers and brewers are sensitive and also the antigen recognized by patients with Crohn's disease. A fraction of S. cerevisiae mannan, Sc500, having high affinity for antibodies in Crohn's patients has been characterized by NMR spectroscopy followed by fragmentation using alkaline elimination, partial acid hydrolysis and acetolysis. The released oligosaccharides were separated by gel filtration on a Biogel P4 column and analyzed by fluorescence labeling, HPLC and methylation analysis. The relationship between structure and antigen activity was measured by competitive ELISA. The antigenic activity of the original high molecular weight mannan could be ascribed to terminal Manα1→3Manα1→2 sequences which are rarely found in human glycoproteins but were over-represented in Sc500 compared to other yeast mannans.
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