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  • Column liquid chromatography  (124)
  • Yeast  (58)
  • wheat  (57)
  • Springer  (239)
  • Cambridge University Press
  • Frontiers Media
  • 2020-2022
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of industrial microbiology and biotechnology 3 (1988), S. 9-14 
    ISSN: 1476-5535
    Keywords: Single cell protein ; Sucrose ; Yeast ; Thermotolerance ; Fermentation ; Kluyveromyces marxianus var.marxianus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Amino acid analyses were undertaken on single cell protein (SCP) produced by thermotolerant strains ofKluyveromyces marxianus var.marxianus grown on sugar cane molasses at 40°C. The maximum conversion of available sugars to biomass at 45°C was only 10.8% (g dry wt.·g−1 total sugars). The amino acid composition of the SCP did not differ markedly from that reported for other yeast species.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Monatshefte für Chemie 116 (1985), S. 1233-1236 
    ISSN: 1434-4475
    Keywords: Stereoselective reduction ; (S)-1-Phenylethanol ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The velocity of reduction of 4-substituted acetophenones by baker's yeast is decreased by electron donating substituents. The steric course, however, is little influenced and (S)-1-arylethanols2 are generally formed with over 90% enantiomeric excess.
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  • 3
    ISSN: 1420-9071
    Keywords: Hydroxamic acid ; cereals ; maize ; wheat ; aphids ; greenbug
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 2-β-Glucosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc), the main hydroxamic acid from maize and wheat, and its aglucone, decreased survival ofSchizaphis graminum reared on artificial diets. Both compounds were toxic for aphids and acted as feeding deterrents, at concentrations as low as 1 mM. The natural concentrations of glucosides of hydroxamic acids in wheat leaves reach up to 6 mmoles/kg fresh weight, thus falling within the range in which DIMBOA-Glc causes delecterious effects to diet-fed aphids.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 46 (1988), S. 47-54 
    ISSN: 1570-7458
    Keywords: Coleoptera ; Curculionidae ; Sitophilus ; S. oryzae ; S. zeamais ; rice weevil ; maize weevil ; geographical strains ; digestion ; amylase ; diet ; barley ; corn ; maize ; rice ; wheat ; amylase inhibitors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé La transformation de rang des valeur d'activité spécifique de l'α-amylase de 4 souches de S. oryzae et de 4 souches de S. zeamais montre que les niveaux les plus élevés de ces enzymes prédominantes s'observent chez les adultes nourris d'orge mondé ou de riz brun á grains longs. Des niveaux intermédiaires d'activité ont été obtenus chez les insectes élevés sur maïs jaune, et les niveaux les plus faibles chez ceux élevés sur blé. Bien que les extraits préparés à partir d'orge présentent une activité inhibitrice de deux isoamylases purifiées de S. oryzae, les niveaux des inhibiteurs naturels α-amylase de ces deux enzymes sont environ respectivement 2,2 et 6,1 fois plus concentrés dans le blé. L'ingestion de ces inhibiteurs d'amylase et la formation d'un complexe enzyme inactive/inhibiteur avec l'amylase secrétée antérieurement, peut rendre compte de la plus faible activité de l'amylase chez les charançons consommant du blé. Le niveau d'amylase de S. oryzae est 2 fois plus élevé que celui de S. zeamais pour toutes les souches élevées sur un régime donné. Des niveaux d'activité significativement différents ont été trouvés suivant les souches pour chacune des deux espèces. Puisque l'amylase est la principale hydrolase digestive de ces espèces, l'intensité de la modification des teneurs en amylase par la consommation de céréales peut indiquer leur adéquation comme hôtes potentiels.
    Notes: Abstract Rank transformation of specific activity values of α-amylase across four strains of Sitophilus oryzae (L.) and four strains of S. zeamais Motschulsky indicates that levels of these predominant enzymes are highest in adults feeding on hulled barley or long-grain brown rice. Intermediate activity levels are found in weevils feeding on yellow corn (maize) and lowest levels are found in wheat-fed weevils. Although extracts prepared from barley contain inhibitory activity against two purified isoamylases from S. oryzae, levels of the naturally-occurring α-amylase inhibitors against these two enzymes are about 2.2-fold and 6.1-fold, respectively, more concentrated in wheat. Ingestion of these amylase inhibitors and formation of an inactive enzyme:inhibitor complex with previously secreted amylase may account for the lower activity of amylase in weevils of both species feeding on wheat. Amylase levels across all strains feeding on a given diet are about 2-fold higher in S. oryzae than in S. zeamais. Significant differences in activity levels were also found between strains in both species. Since α-amylase is a predominant digestive hydrolase in these species, the degree to which cereal diets affect amylase levels may indicate their suitability as potential hosts.
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  • 5
    ISSN: 1432-0983
    Keywords: Ribosomal protein ; Immunological homology ; Yeast ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Polyclonal antibodies raised against ribosomal protein (r-protein) L2 of Schizosaccharomyces pombe were used to check for cross-reaktions with total r-proteins of rat liver. Using this procedure, the rat liver r-proteins, L4 and L24, were identified as being immunologically related to yeast L2. In addtional, homologies between rat liver L4 and L24 were detected. The possible implications for the regulation of r-protein synthesis are discussed.
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  • 6
    Electronic Resource
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    Springer
    Current genetics 13 (1988), S. 291-297 
    ISSN: 1432-0983
    Keywords: Yeast ; Nuclear matrix ; Plasmid stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Acentric yeast plasmids are mitotically unstable, apparently because they cannot freely diffuse after replicating and therefore are not included in the daughter nucleus. This behavior could result if plasmids remain attached to structural elements of the nucleus after replicating. Since DNA replication is believed to take place on the nuclear matrix, we tested whether there was a correlation between the mitotic stability of a given plasmid and the extent to which it was found associated with residual nuclear structures. Residual nuclei were prepared from yeast nuclei by extraction with either high salt, 2 M NaCl, or low salt, 10 mM lithium diiodosalicylate (LIS). Hybridization analysis was used to estimate the fraction of plasmid molecules remaining after nuclei were extracted. We examined the extent of matrix association of three ARSI plasmids, Trpl-RI circle (1.45 kb), YRp7 (5.7 kb) and pXBAT (45.1 kb) with mitotic loss rates ranging from 3–25%. In addition we examined the matrix binding of the endogenous 2 μm plasmid and the 2 μm-derived YEp 13 which is relatively stable in the presence of 2 μm and less stable in cir° strains. Among the ARS1 plasmids we observed a negative correlation between stability and matrix association, consistent with models in which binding to the nuclear matrix prevents passive segregation of ARS1 plasmid molecules. No such correlation was observed among the 2 μn plasmids. Among all plasmids examined there is a positive correlation between size and matrix association.
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  • 7
    ISSN: 1432-0983
    Keywords: Yeast ; Cloning ; ODC ; Complementation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A DNA fragment containing the gene encoding orotidine 5′-phosphate decarboxylase (ODC) from the yeast, Schwanniomyces occidentalis (formerly castellii) has been isolated from a genomic library constructed in the S. cerevisiae expression vector, pYcDE8. A recombinant plasmid, p2-lA, containing a 2.47 kb insert was shown to complement the ura3-52 mutation of several strains of S. cerevisiae. This DNA insert was shown to be from Schwanniomyces occidentalis by Southern hybridization analysis. A restriction enzyme cleavage map of the insert has been derived and the ODC gene localized to a 1.1 kb region by deletion analysis. In addition, we have demonstrated that expression of ODC is not dependent on the ADHI promoter carried on pYcDE8. This is the first report of the cloning of a gene from a member of the genus Schwanniomyces.
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  • 8
    ISSN: 1432-0983
    Keywords: Yeast ; Gene regulation ; Saccharomyces cerevisiae ; PDCI promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 870 by promoter fragment of the PDC1 gene that includes the carbon source dependent regulatory regions was investigated using 5′ and 3′ promoter deletions. The results indicate that glucose and ethanol regulation of PDC1 transcription are independently controlled by distinct cis-acting regions. The consensus sequence AAATCGATA may play a role in this regulation, while the sequence (ATCA)AACCT may be important in transcription initiation.
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  • 9
    ISSN: 1432-0983
    Keywords: Gene regulation ; Cell cycle ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Expression of the thymidylate synthase gene (TMPI) of Saccharomyces cerevisiae increases during the late G1 phase of the cell cycle. Using a series of gene fusions, which have placed the Escherichia coli lacZ gene under transcriptional and translational control of different portions of the TMPI gene, we have demonstrated the existence of three different regions which are important for expression. One of these regions, which was localized to within 270 base pairs of the translation start codon, is involved in the periodic expression of TMPI transcript. A second region, the deletion of which resulted in reduced levels of TMPI expression, is at least partially encoded by DNA sequences between 270 and 377 base pairs upstream of the translation start codon. A third region, located within the N-terminal 112 codons of the TMPI gene, apparently encodes information involved in a post-translational control mechanism.
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  • 10
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    Current genetics 10 (1985), S. 87-93 
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; oxi2 mutations ; Functional suppressors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A semidominant nuclear suppressor, callednam6, ofoxi2-V276 mitochondrial mutation has been isolated and characterized. The nuclear character ofnam6 was proved by its retention inrho° strains, lack of mitotic segregation in diploids and meiotic 2:2 segregation in tetrads. The specificity ofnam6 was tested on 315mit − mutations of four mitochondrial genes (oxi1, oxi2, oxi3, andcob-box). It suppresses clearly only three mutations in theoxi2 gene, restoring partially or completely cytochrome aa3 formation. The results suggest a functional character of the suppression.
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  • 11
    ISSN: 1432-0983
    Keywords: Sporulation ; Yeast ; Transcription ; Meiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have characterized 46 hybrid phage which hybridize preferentially to mRNA from sporulating cells. Cross-hybridization experiments demonstrate that 27 distinct SPR (Sporulation regulated) sequences are represented among these phage. The SPR genes can be grouped into three classes: early, middle, and late. The early class shows an accumulation of transcripts soon after transfer to sporulation medium and continues to accumulate RNA throughout sporulation. Transcripts of the middle class increase in level at about the time of DNA synthesis, rise rapidly in abundance until meiosis II, then accumulate more slowly for at least the next 3 h. Late gene transcripts begin to accumulate at about the time of meiosis I, increase 10- to 20-fold in the next 2 h, then remain constant in late sporulating cells.
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  • 12
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    Current genetics 13 (1988), S. 455-460 
    ISSN: 1432-0983
    Keywords: Yeast ; Gene conversion and mutation ; CDC8 locus ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The induction of mitotic recombination in theCDC8 locus was studied in a diploid strain heteroallelic forcdc8 mutations (cdc8-1/cdc8-3); mitotic reversion was studied in strainscdc8-1/cdc8-1 andcdc8-3/cdc8-3. Conversion and reversion did not occur in those cells blocked at the S stage of the cell cycle by exposure to a nonpermissive temperature. In stationary phase cells irradiated just prior to exposure to temperature stress, the induction of recombinants was rather low and the induction of revertants was minimal. Conversely, a significant induction ofcdc + occurred in logarithmic phase cells subjected to the same treatment. Irradiation of synchronously dividing cultures revealed that intragenic recombination occurs at all three stages of the cell cycle- G1, S and G2. It was also found that UV-induced gene reversion can occur during the S and G2 stages, but not during the G1 stage of the cell cycle.
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  • 13
    ISSN: 1432-0983
    Keywords: Posttranslational processing ; Ribosomal protein gene ; Transcript mapping ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Analysis of the primary structure of the gene for yeast ribosomal protein S31 revealed two unusual features. First, an intron of 312 nucleotides is located within the 5′-untranslated region. Second, the coding sequence for the known amino-terminal peptide of the protein starts 13 codons downstream of the ATG initiation codon, suggesting that S31 is synthesized as a precursor which undergoes post-translational processing to the mature protein. Primer extension analysis showed that transcription of the S31 gene starts at multiple sites. The 5′-flanking region of the gene contains several, previously described, conserved sequence elements that may play a role in the coordinate expression of yeast ribosomal protein genes.
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  • 14
    ISSN: 1432-0983
    Keywords: DNA polymerase ; Yeast ; Immunoscreening ; Cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Partially overlapping fragments of the gene encoding yeast DNA polymerase I have been cloned by immunological screening of a yeast genomic library constructed in the phage λ expression vector λgt11. The three gene fragments we analyzed in detail encode part of a yeast protein that has been identified as yeast DNA polymerase I, because it shares with this enzyme a number of antigenic determinants. In fact, the yeast protein fragments expressed by the recombinant phages react with both polyclonal and monoclonal antibodies raised against different, highly purified preparations of DNA polymerase I. Moreover, they can be used to affinity purify antibodies specifically reacting with active DNA polymerase I polypeptides and they compete with the yeast enzyme for binding to antibodies that inhibit catalytic activity. The gene is located on chromosome XIV in the yeast genome, and it is transcribed as a 5.2 kb mRNA.
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  • 15
    ISSN: 1432-0983
    Keywords: Mitochondria ; Yeast ; Deletions ; RNA stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two cob − deletion mutants are characterized. One of them, M9410, is deleted for 911 by of the noncoding sequences only which separate tRNAGlu and cob exon 1; it thus lacks most of the sequence encoding the 957 by long cob leader (Bonitz et al. 1982) and some 20 by 5′ to it. The end points of this deletion coincide with 31 by long direct repeats in wild type mtDNA. The other mutant, M9391, is deleted for all cob coding sequences and most of the cob leader sequence but it retains the 5′ terminal 261 by of this leader. Northern analysis revealed that M9410 totally lacks cob mRNA or pre-mRNA. The large deletion M9391 in contrast accumulates a 13S RNA which probably results from transcription through the junction, which ligates sequences of the cob leader to sequences of the cob-oli1 intergenic spacer.
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  • 16
    ISSN: 1432-0983
    Keywords: Mitotic recombination ; DNA repair ; Yeast ; RAD52
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The RAD52 gene is required for much of the recombination that occurs in Saccharomyces cerevisiae. One of the two commonly utilized mutant alleles, rad52-2, increases rather than reduces mitotic recombination, yet in other respects appears to be a typical rad52 mutant allele. This raises the question as to whether RAD52 is really necessary for mitotic recombination. Analysis of a deletion/insertion allele created in vitro indicates that the null mutant phenotype is indeed a deficiency in mitotic recombination, especially in gene conversion. The data also indicate that RAD52 is required for crossing-over between at least some chromosomes. Finally, examination of the behavior of a replicating plasmid in rad52-1 strains indicates that the frequency of plasmid integration is substantially reduced from that in wild type, a conclusion consistent with a role for RAD52 in reciprocal crossing-over. Analysis of recombinants arising in rad52-2 strains suggests that this allele may result in the increased activity of a RAD52-independent recombinational pathway.
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  • 17
    ISSN: 1432-0983
    Keywords: Informational suppressors ; Modifier ; Yeast ; tRNAs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants of Saccharomyces cerevisiae were selected that would interact with ochre (UAA) suppressors so as to allow ochre -suppressor dependant amber (UAG) suppression, but which do not exhibit opal (UGA) suppression. Strains mutant at four distinct loci were isolated, and two of these are recessive mutations while the other two behave as dominants or semidominants. MOS3 has some suppressor activity in the absence of a resident SUP4-o gene and shares other characteristics with previously described omnipotent suppressors. MOS4, mos1 and mos2, on the other hand, exhibit no suppressor activity in the absence of a resident SUP4-o gene but do exhibit suppression of UAG alleles when there is a resident SUP4-o gene. These latter modifier strains do not interact with a SUP4-o gene to suppress UGA alleles. By genetic and physiological criteria the MOS4, mosl, and most mutations appear to be different than previously described allosuppressors or modifiers of suppression.
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  • 18
    ISSN: 1432-0983
    Keywords: Yeast ; CDC8 gene ; DNA replication, recombination, mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The conditional cdc8 mutant is known to be defective, under restrictive conditions, in the elongation of DNA during synthesis. In yeast the CDC8 gene encodes thymidylate kinase. We show here that UV-induced gene conversion and gene mutation events require the participation of this CDC8 gene. Thus, the same thymidylate kinase is incolved both in DNA replication and in UV-induced gene conversion and gene mutation in yeast.
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  • 19
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    Mycopathologia 102 (1988), S. 3-8 
    ISSN: 1573-0832
    Keywords: Yeast ; carbon assimilation profiles ; liquid medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Liquid medium assays for yeasts carbon assimilation tests are the more precise but longer methods. For rapid and automated yeasts identification purposes we analysed the assimilation of 34 carbon compounds by 149 reference strains. Assays were carried in liquid shaken medium (Autobac∘ system) and readings were nephelemetric. Valuable results are obtained in 72 hours and their analysis allowed us to classify substrates for their ability to minimize the number of doubtful results.
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  • 20
    ISSN: 1573-0832
    Keywords: odors ; volatiles ; microflora ; fungi ; wheat ; seasonality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The fungal odor compounds 3-methyl-l-butanol, l-octen-3-ol and 3-octanone were monitored in nine experimental bins in Winnipeg, Manitoba containing a hard red spring wheat during the autumn, winter and summer seasons of 1984–85. Quality changes were associated with seed-borne microflora and moisture content in both ventilated and non-ventilated bins containing wheat of 15.6 and 18.2% initial moisture content. All three odor compounds occurred in considerably greater amounts in bulk wheat in non-ventilated than in ventilated bins, particularly in those with wheat having 18.2% moisture content. The presence of these compounds usually coincided with infection of the seeds by the fungi Alternaria alternata (Fr.) Keissler, Aspergillus repens DeBarry, A. versicolor (Vuill.) Tiraboschi, Penicillium crustosum Thom, P. oxalicum Currie and Thom, P. aurantiogriseum Dierckx, and P. citrinum Thom. High production of all three odor compounds in damp wheat stored in non-ventilated bins was associated with heavy fungal infection of the seeds and reduction in seed germinability. High initial moisture content of the harvested grain accelerated the production of all three fungal volatiles in non-ventilated bins.
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  • 21
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    Nutrient cycling in agroecosystems 17 (1988), S. 147-151 
    ISSN: 1573-0867
    Keywords: time of zinc application ; wheat ; zinc sulphate ; zinc oxide ; zinc uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field experiments with wheat were conducted for two years on flood plain alluvial soils to study the effectiveness of soil application of zinc sulphate and zinc oxide at 0, 15, 45, 60, 75 and 90 days after sowing. Yield and zinc uptake of wheat increased significantly with the application of zinc. Delaying the application of both zinc sulphate and zinc oxide up to 45 days of sowing did not adversly affect the zinc nutrition of wheat. However, delaying the application for 75 or 90 days after sowing eliminated the response. Zinc sulphate, when applied within 60 days of sowing performed better than zinc oxide. In a laboratory study, zinc sulphate maintained a higher level of zinc in the soil solution than zinc oxide at least over a 3-week period.
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  • 22
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    Nutrient cycling in agroecosystems 17 (1988), S. 137-146 
    ISSN: 1573-0867
    Keywords: Ammonium-nitrate-potassium interaction ; accumulation of reduced nitrogen ; wheat ; nitrification inhibitor ; grain ; stover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A greenhouse experiment with wheat in 3L pots filled with a sandy loam soil in a factorial design was conducted to determine the effect of potassium on nitrogen utilization. Nitrogen was applied in three NH4-N/NO3-N ratios, 0/100, 25/75 and 50/50, at three levels: 0.75, 1.50 and 3.00gN/pot, and potassium was applied at three levels: 0, 0.5 and 1.0gK/pot. The higher levels of nitrate nitrogen with or without potassium reduced dry matter yields drastically, while the same levels of a NH4-N/NO3-N mixture of 50/50 with applied potassium reduced yields only slightly. Highest grain yield and total yield were obtained with a 25/75 mixture of ammonium/nitrate nitrogen with added potassium. Potassium addition to soil increased the utilization of nitrogen fertilizers, particularly when the ratio of ammonium to nitrate was increased. The highest uptake of reduced nitrogen was at the highest level of the ammonium to nitrate nitrogen ratio (50/50) when potassium was applied. Tillering was enhanced by an increased ammonium ratio in the nitrogen mixture, and by potassium.
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  • 23
    ISSN: 1573-0867
    Keywords: farmyard manure ; maize ; nitrogen ; phosphorus ; rice ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field experiments with rice-wheat rotation were conducted during five consecutive years on a coarse-textured low organic matter soil. By amending the soil with 12t FYM ha−1, the yield of wetland rice in the absence of fertilizers was increased by 32 per cent. Application of 80 kg N ha−1 as urea could increase the grain yield of rice equivalent to 120 kg N ha−1 on the unamended soil. Although the soil under test was low in Olsen's P, rice did not respond to the application of phosphorus on both amended and unamended soils. For producing equivalent grain yield, fertilizer requirement of maize grown on soils amended with 6 and 12 t FYM ha−1 could be reduced, respectively to 50 and 25 per cent of the dose recommended for unamended soil (120 kg N + 26.2 kg P + 25 kg K ha−1). Grain yield of wheat grown after rice on soils amended with FYM was significantly higher than that obtained on unamended soil. In contrast, grain yield of wheat which followed maize did not differ significantly on amended or unamended soils.
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  • 24
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    Nutrient cycling in agroecosystems 6 (1985), S. 171-176 
    ISSN: 1573-0867
    Keywords: Chloride ; phosphorus ; salinity ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A pot study with four levels each P (control, 6.5, 13.0, 19.5 mg kg−1) and Cl− in irrigation water (control, 30, 60, 90me l−1) was carried out to test Cl− and P interaction on wheat. It has been found that optimum P nutrition alleviates the toxic effect of excess Cl−. Highest P rate resulted in a significant yield increase upto a Cl− level of 60me l−1. Cl− depressed P content in the plant only at a Cl− level of 90me l−1, while P rates had no major impact on the Cl− concentration in the plant. Lower Cl− concentration at higher P rates are rather a dilution effect than an antagonistic one. With optimum P supply higher Cl− contents in the plants were tolerated than with a low P supply.
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  • 25
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    Nutrient cycling in agroecosystems 8 (1985), S. 91-96 
    ISSN: 1573-0867
    Keywords: chickpea ; Egyptian clover ; field experiments ; K uptake ; mustard ; wheat ; yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A field experiment on a potassium deficient Typic ustochrept soil of the University Farm at Pura, Kanpur compared the responses of wheat, chickpea, mustard and Egyptian clover to added potassium for two consecutive years commencing in 1979–80. All the crops responded to potassium fertilization. Sensitivity of various crops to potassium deficiency was in the order: Egyptian clover 〉 chickpea 〉 wheat 〉 mustard. Higher sensitivity of Egyptian clover and chickpea compared to wheat was in agreement with their comparatively high root cation exchange capacity. Mustard by contrast showed higher root cation exchange capacity and exhibited least potassium sensitivity because of its higher potassium utilization efficiency. Leaf K content and total uptake of potassium by all crops was increased by applying potassium upto the highest level.
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  • 26
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    Nutrient cycling in agroecosystems 8 (1985), S. 173-178 
    ISSN: 1573-0867
    Keywords: critical limit of Mn ; highly permeable soils ; methods and rates of Mn ; Mn deficiency ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two field experiments were conducted on Mn-deficient soils to evaluate the efficiency of rates, methods and time of MnSO4.H2 O application for wheat. Manganese sulphate was broadcast and mixed in soils at the rate of 5 to 50kg Mn ha−1 before seeding and 10 to 40 kg Mn ha−1 as top dress at 28 days — just before first irrigation. Three sprays of 1% MnSO4·H2O unneutralised solutions were applied, the first at 26 days — 2 days before first irrigation and the others afterward at weekly intervals. Both the methods caused a significant and marked increase in grain yield. Three foliar sprays were as effective as soil applications of 20 to 40 Kg Mn ha−1 before seeding. The difference in grain yield resulting from soil applications of Mn before seeding and applications at the first irrigation was not significant. The DTPA-Mn status of 20 fields, selected on the basis of varying degree of Mn deficiency, was related to grain yield (r = 0.77**). Also grain yield of all the experiments had a significant correlation with Mn content of grain (r = 0.55** to 0.82**) and straw (r = 0.77** to 0.82**). The critical limits calculated by statistical method were 1.25, 2.18 and 3.5 mg Mn kg−1 soil for severe deficiency, deficiency and latent deficiency respectively for wheat.
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  • 27
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    Nutrient cycling in agroecosystems 17 (1988), S. 165-176 
    ISSN: 1573-0867
    Keywords: Low-grade phosphate rock ; acid extracts ; P fertiliser ; available P ; wheat ; soil pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Low grade phosphate rock (PR), containing high amounts of oxides of iron and aluminium is neither suitable for fertiliser production nor useful for direct application to annual crops. The fertiliser effectiveness of P extracted by H2SO4 from a low-grade phosphate rock, PR (Christmas Island C-grade PR) was evaluated for wheat (Triticum aestivum L.) on a calcareous loam (pH 8.4) and a non-calcareous loam (pH 6.9) in field and glasshouse experiments. Superphosphate was used to compare the performance of the acid extracts of PR. In the non-calcareous loam soil, crop establishment and yield were significantly reduced by the acid extracts of PR due to increased acidity. In the calcareous soil, however, the acid extracts of PR performed as well as superphosphate; similar or even higher crop yields were obtained with the former, especially when applied near the seed. The acid extracts of low-grade PR may, therefore, have a role in calcareous soils, where the extract can be applied directly or added in the irrigation waters to supply P to crops.
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    Nutrient cycling in agroecosystems 16 (1988), S. 137-155 
    ISSN: 1573-0867
    Keywords: Superphosphate ; rock phosphate ; wheat ; oats ; barley ; field experiment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Nine soil tests for phosphate were evaluated for predicting the yield and P content of wheat, barley and oats grown on a sandy soil in Western Australia: Olsen, modified Olsen 1 (soil:solution ratio 1:5), modified Olsen 2 (soil:solution ratio 1:50), Colwell, Bray 1, Bray 2, modified Bray 2T (shaking time 10 minutes), modified Bray 2C (pH 3.7) and lactate. The soil had been fertilized 5 years previously with 20 levels each of superphosphate (OSP, range 0 to 400 kg P ha−1) and Queensland rock phosphate (QRP, range 0 to 20 000 kg P ha−1). For each species and fertilizer taken separately, all the tests, except for lactate, gave a good prediction of yield. When data for OSP and QRP were pooled, Bray 2 and modified Bray 2T tests were unsatisfactory predictors of both yield and P content. A linear relationship (P 〈 0.05) between mean soil tests value (χ) and the standard deviation (σ χ ) of the test value was observed for each soil test. For QRP, the results for lactate were the most variable (i.e.σχ/χ was greatest) followed by modified Olsen 2 〉 Bray 1 〉 Bray 2 〉 Olsen 〉 modified Bray 2C 〉 modified Olsen 1 〉 modified Bray 2T 〉 Colwell. The order for OSP fertilized soil was Bray 1 〉 modified Bray 2T 〉 Bray 2 〉 Olsen 〉 Colwell 〉 modified Bray 2C 〉 modified Olsen 1 〉 lactate 〉 modified Olsen 2. For combined OSP and QRP data, the results of the Olsen 1 and Colwell extractions were the least variable. Errors in the prediction of yield (σ Y ) for all crops resulting from an error in soil test values (σχ) were calculated. For OSP-fertilized soil variability in values for the Bray-1 test provided the highest error (about 16%) in the prediction of the yield, followed by Bray 2 (12%) 〉 Bray 2T (10%) 〉 Olsen (8%) 〉 Colwell (7%) 〉 modified Bray 2C (6%) 〉 lactate (4%). Maximum error was at yields of about 65% of maximum yield. For soil fertilized with QRP, lactate provided the highest error (about 10%) in the prediction of yield, followed by the other tests (〈 6%). Maximum error was at yields of about 35% of maximum yield. The Colwell soil test gave the most accurate overall prediction of yield for both fertilizers.
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  • 29
    ISSN: 1432-0983
    Keywords: Yeast ; Carbon catabolite repression ; Oncogene-related genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The “start” cell division control genes CDC36 and CDC28 have been reported to contain a certain sequence homology to tissue oncogenes (ets and some protein kinase encoding oncogenes respectively). Here we report that temperature sensitive mutations in these genes are suppressed in cytoplasmic “petite” mutants and catabolite repression resistant mutants.
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  • 30
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    Current genetics 10 (1985), S. 253-260 
    ISSN: 1432-0983
    Keywords: Yeast ; RNA polymerase I ; Promoter ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Deletions in the promoter region of the 37S pre-rRNA operon in yeast were constructed and analysed in vivo using an artificial ribosomal minigene present on an extrachromosomal yeast vector. Sequences required for correct transcription initiation were found to be located between positions −192 and +15 relative to the start; a 5′-deletion down to position −133 reduces the transcription yield of the minigene at least five-fold. To allow detection of transcription of the minigene in isolated nuclei of yeast transformed with a minigene-bearing plasmid we attempted to increase the minigene copy number. The transcription yield in vivo appeared not to be proportional to the copy number but was found to be greatly enhanced when two or three mini-genes are present in tandem. α-Amanitin sensitivity of transcription of these minigenes in isolated nuclei proved that RNA polymerase I is responsible for their transcription.
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  • 31
    ISSN: 1432-0983
    Keywords: Yeast ; DNA repair mutants ; Allelism test ; Psoralen plus UVA
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    Notes: Summary In the yeast Saccharomyces cerevisiae, allelism between the psol-1 and the rev3-1 mutants on the one hand and the pso2-1 and snm1 mutants on the other, is demonstrated by the comparison of phenotypes, complementation tests and meiotic segregation analysis.
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  • 32
    ISSN: 1432-0983
    Keywords: Fusion ; Protoplast ; Saccharomyces ; Yeast
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Protoplasts of Saccharomyces cerevisiae his1 trp2 resistant to acriflavine and able to ferment galactose and of Saccharomyces fennentati arg resistant to DL-p-fluorophenylalanine and able to ferment lactose were fused. As a result of fusion two types of prototrophic hybrids were obtained. Type 1 hybrids were able to grow on medium with galactose or lactose as sole carbon source and were sensitive to acriflavine and resistant to DL-p-pfluorophenylalanine. Type 2 hybrids were able to grow on medium with galactose as sole carbon source and were resistant to acriflavine and sensitive to DL-p-fluorophenylalanine.
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  • 33
    ISSN: 1432-0983
    Keywords: Yeast ; FLP-FRT ; BFBC ; Gene conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A YEp chimaeric plasmid containing URA3 and SMR1 [sulfometuron methyl resistant (SMR) allele of ILV2] as selectable markers, and the 2 μm site-specific recombination FLP recognition target (FRT), was integrated at the ilv2-Δ1 site in chromosome XIII in a cir°] haploid. Southern analysis defined two integrant structures. Structure I had URA3 distal and SMR1 proximal to FRT whereas in structure II both markers were distal to FRT. Selectable markers were stably inherited in [cir°] haploids and [cir°] diploids heterozygous for the integrant and ILV2. Approximately 14% of heterozygous [cir +] diploid cells exhibited homozygotization for the distal (500 kb) ade4 marker in trans. In [cir +] diploids FLP-FRT recombination resulted in the simultaneous loss of both structure II markers, whereas the structure I distal URA3 marker loss always preceded the variable loss of the proximal SMR1 marker. URA− cells continued to segregate for loss of SMR1 until stable URA− SMR or URA−SMS cells were produced. Gene conversion was identified in stable URA−SMR cells that were homozygous SMR1/SMR1 but contained wild type ILV2 restriction endonuclease sites. These observations support a model based on concerted FLP-FRT action resulting from the secondary integration of native 2 μm DNA followed by unequal sister chromatid exchange (USCE) within inverted FRTs. The resultant chromatid bridge resulted in a double-stand break. Fusion of the broken ends of sister chromatids generated a breakage-fusion-bridge cycle (BFBC). Repeated rounds of the BFBC resulted in proximal marker loss and the generation of additional double-strand breaks. Recombinogenic properties of the double-strand break initiated events leading to homozygotization and gene conversion.
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  • 34
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    Current genetics 9 (1985), S. 279-284 
    ISSN: 1432-0983
    Keywords: Virus-like particles ; Double-stranded RNA ; Yeast ; Yarrowia lipolytica
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    Topics: Biology
    Notes: Summary Four out of the 24 strains of the yeast Yarrowia lipolytica we have checked for the presence of virus-like particles (VLPs) proved to contain encapsidated double-stranded RNA (dsRNA) molecules, 4.9 kb long. A major VLP polypeptide of MW 80,000 was observed in all 4 cases, and a second one of MW 77,000 in three cases. dsRNA from the VLPs harboring only the larger polypeptide showed little homology with the 3 others. We have found no homology between VLP dsRNAs and host DNA or dsRNAs from Saccharomyces cerevisiae, and no relationship between the presence of VLPs and a possible killer phenomenon in Y. lipolytica.
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  • 35
    ISSN: 1432-0983
    Keywords: D-xylose fermentation ; Yeast ; Protoplast fusion ; Ploidy
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    Topics: Biology
    Notes: Summary Protoplast fusion technique was employed in the preparation of presumptive diploid, triploid and tetraploid strains of the D-xylose fermenting yeast C. shehatae CBS2779. Prototrophic selection technique was employed for the recovery of presumed fusant strains. The hybrid nature of the presumptive diploid, triploid and tetraploid strains was confirmed by analysing I) the nuclear condition; II) the cell size and the cell volume of the parental and fusant strain; III) the cellular DNA content and IV) the induced and spontanenous mitotic segregation of properties in these strains. The increased level of ploidy was found to have an effect on the rate of ethanol production from D-xylose.
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  • 36
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    Current genetics 9 (1985), S. 533-538 
    ISSN: 1432-0983
    Keywords: cDNA hybridisation ; UV inducible RNA ; Yeast
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    Topics: Biology
    Notes: Summary Differential colony hybridisation has been used to identify DNA sequences in Saccharomyces cerevisiae corresponding to RNA transcripts whose levels increase 5–10 fold following UV-irradiation. Four sequences have been identified, three of which share sequence homology and hybridise to the same set of genomic DNA fragments. The fourth sequence appears to be distinct, however each DNA sequence hybridises to a similar sized RNA transcript which is approximately 4.0 kb long. The relationships between these DNA sequences and their potential protein products is discussed.
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  • 37
    ISSN: 1432-0983
    Keywords: Yeast ; TEF genes ; Gene disruption
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    Notes: Summary Two genes,TEF1 andTEF2, encode the protein elongation factor EF-1α in the yeastSaccharomyces cerevisiae. We have generated yeast haploid strains containing eitherTEF1 orTEF2 interrupted by insertion of a large piece of foreign DNA. Cells which contain either one functional copy of the EF-1α genes are viable. In contrast, attempts to isolate a yeast haploid strain with bothTEF1 andTEF2 inactivated have failed suggesting that the double gene disruption is a lethal event.
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  • 38
    ISSN: 1432-0983
    Keywords: Yeast ; cdc8-1 mutation ; Mitotic recombination
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    Notes: Summary In a diploid strain homozygous for the cdc8-1 mutation, a block in DNA synthesis caused by restrictive temperature resulted in a significant increase in the frequency of intragenic recombination at the HOM2 locus. Under restrictive conditions, incorporation of radioactivity into DNA was reduced to 2% of the control and alkaline sucrose gradient centrifugation revealed that only short DNA fragments were synthesized. There was no considerable fragmentation of template DNA during incubation of cdc8-1 strains under restrictive conditions.
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  • 39
    ISSN: 1432-0983
    Keywords: Telomeres ; Recombination ; Yeast
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    Topics: Biology
    Notes: Summary Natural termini from macronuclear DNA of the ciliated protozoans Tetrahymena thermophila and Oxytricha fallax can support telomere formation in yeast. However, plasmids carrying these ciliate termini are modified by the addition of DNA which hybridizes to the synthetic oligonucleotide poly [d(C-A)], a sequence which also hybridizes to terminal restriction fragments from yeast chromosomes but not to Tetrahymena or Oxytricha macronuclear DNAs. Thus, in yeast, the creation of new telomeres on ciliate termini involves the acquisition of yeast-specific terminal sequences presumably by either recombination or non-templated DNA synthesis. The RAD52 gene is required for the majority of yeast mitotic and meiotic recombination events. Moreover, the absence of an active RAD52 gene product results in high rates of chromosome loss. Here we demonstrate that terminal restriction fragments from Tetrahymena macronuclear ribosomal DNA (rDNA) support the formation of modified telomeres in a yeast strain carrying a defect in the RAD52 gene. Moreover, linear plasmids bearing these modified ciliate termini are stably propagated in rad52 − cells.
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  • 40
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    Current genetics 9 (1985), S. 259-262 
    ISSN: 1432-0983
    Keywords: Yeast ; Ty1 ; Trans ; Deletion
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    Topics: Biology
    Notes: Summary DEL1 strains of the yeast Saccharomyces cerevisiae exhibit a high rate of deletions of the three linked genes, CYC1, OSM1, and RAD7. Classical genetic methods showed that DELI segregated as a single Mendelian gene closely linked to CYC1. In addition, genetic evidence suggested that DEL1 was both cis- and trans-dominant (Liebman et al. 1979). Molecular analysis of deletions isolated from a haploid DEL1 strain established that deletion formation was mediated by recombination between yeast transposable elements, Ty's (Liebman et al. 1981). We now report the molecular characterization of deletions isolated from diploids in the trans configuration. This analysis reveals that these deletions probably arose in a two-step process involving mitotic recombination followed by Ty-mediated deletion formation in cis.
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  • 41
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    Current genetics 9 (1985), S. 285-291 
    ISSN: 1432-0983
    Keywords: Killer toxin ; Plasmid selection ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transformants of sensitive yeast strains containing an expressed cDNA copy of the yeast killer toxin-immunity gene could be selected for by exposure to added killer toxin. For strain AH-22 the transformation frequency was approximately 10% that obtained by selection for leucine prototrophy. The procedure required time for expression of immunity prior to selection, and a screening step to remove non-transformed survivors. Under conditions where active toxin was produced, transformants containing the toxin-immunity gene were at a selective advantage, and cells losing the plasmid were killed. This resulted in self selection of transformants, and affords a way of maintaining plasmid stability in protrophic strains.
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  • 42
    ISSN: 1432-0983
    Keywords: Yeast ; Ribosomal protein gene ; Sequence analysis ; Conserved elements
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    Notes: Summary Computer analysis has previously revealed the presence of a 12-nucleotide common sequence element (AACATC CA TG T A G CA; HOMOL1) in the upstream regions of several yeast ribosomal protein genes. By extending the sequence analysis of the 5′-flanking regions of a number of other ribosomal protein genes (including those encoding S10-1, S10-2, S33 and L16-2) we could establish that HOMOL1 occurs upstream of most but not all yeast ribosomal protein genes. Apart from HOMOL1 an additional conserved sequence (ACCCATACATT A T ; RPG-box) was detected in front of nearly all yeast ribosomal protein genes, although in some cases it is present in the opposite orientation in the other strand. There seems to be no correlation between the occurrence of one box and that of the other. However when both boxes are present the RPG-box is always located 3′ to the HOMOL1-sequence mostly at a distance of only a few nucleotides. A further one-to-one comparison of the upstream regions of several yeast ribosomal protein genes revealed extensive additional sequence homologies that are suggested to be involved in the coordinate control of ribosomal protein gene expression in yeast.
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  • 43
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    Current genetics 9 (1985), S. 529-532 
    ISSN: 1432-0983
    Keywords: Yeast ; UV-inducible proteins ; rad mutants
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    Topics: Biology
    Notes: Summary Two UV-inducible proteins have been detected in the yeast, Saccharomyces cerevisiae. The proteins have molecular weights of 78,000 Daltons and 23,000 Daltons. This induction is specific for UV-irradiation as exposure to X-rays, mitomycin C and heat shock does not result in the synthesis of the proteins. The larger (78 kD) protein is induced in various rad strains and in a π° cir° strain. Attempts are being made to isolate the genes coding for these inducible proteins.
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  • 44
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    Current genetics 9 (1985), S. 653-660 
    ISSN: 1432-0983
    Keywords: Gene expression ; Yeast ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Efficient expression of theEscherichia coli ZeuB (ß-isopropylmalate dehydrogenase) gene occured in yeast after in vitro DNase digestion and religation of plasmid bound ZeuB and the yeastIIIS3 DNA which placed the 5′ end of the yeastHIS3 gene immediately adjacent to the coding region of theE. coli leuB gene. Two structurally distinct classes of gene fusions were constructed, each involved portions of the yeastHIS3 gene which contributed DNA sequences responsible forleuB expression in yeast. The first class involved fusion of theHIS3 coding region to bacterial DNA resulting in the production of a fusion protein with ß-isopropylmalate dehydrogenase activity. The second class consisted of bacterial DNA, including theleuB coding region, fused to theHIS3 promotor region with the absence of any portion of theIIIS3 coding region. In both constructions theIIIS3 promotor region is required for transcription, however, translation of the class two fusion is initiated at a bacterial DNA coded AUG, and the 5′ end of the mRNA coded by theleuB gene mapped predominantly at bacterial DNA sequences. The DNA sequence responsible for the 5′ end of theHIS3 mRNAs remain in the class two gene fusions but this did not preclude the initiation of transcription at bacterial DNA sequences. The pattern of mRNA initiation at bacterial DNA suggests that DNA sequences at, or adjacent to, the site of transcription initiation are involved in the determination of the sites of initiation, and perhaps the frequency at which initiation occurs.
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  • 45
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    Current genetics 9 (1985), S. 147-155 
    ISSN: 1432-0983
    Keywords: Killer ; Yeast ; Linear plasmid ; Sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Some strains of the yeast Kluyveromyces lactis contain a pair of linear DNA plasmids, k1 and k2, 8.8 and 13.8 kilobase pairs long, respectively. Simultaneous presence of the two plasmids confer a killer phenotype on the cell by producing a toxin which blocks the growth of sensitive yeast species. Previous genetic studies have suggested that the toxin protein is coded by the k1 plasmid. We have now determined the total nucleotide sequence of k1 DNA. The genome is 8,874 base pairs in length. It contains four protein-coding reading frames, three transcribed from one strand and the fourth transcribed from the complementary strand and has terminal inverted repeats of 202 base pairs. Nuclease S1 mapping confirmed this arrangement and showed that these genes are transcribed. The terminal repeats and the four genes form an extremely compact genome, with some overlapping of genes. All four genes use highly biased codons, 86% of them having A or T at the wobble position, reminiscent of yeast mitochondrial genes. Three genes share a very similar 5′ leader sequence. The nature of gene products is discussed in the light of what is known of the excreted toxin protein.
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  • 46
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    Current genetics 9 (1985), S. 179-181 
    ISSN: 1432-0983
    Keywords: Blasticidin S ; Yeast ; Resistant mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Blasticidin S-resistant mutants of S. cerevisiae were isolated and characterized. Resistant mutations were found to fall into two complementation groups. A single recessive nuclear gene was responsible for each group, donated as bls1 and bls2, respectively. A gene bls1 was linked to an ilv3 gene located on the right arm of chromosome X. The resistant phenotypes from both genes were not associated with ribosomes known to be target sites of Blasticidin S, when analyzed by poly(U)-directed polyphenylalanine synthesis. The resistant mechanisms of the mutations are discussed in this paper.
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  • 47
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    Current genetics 9 (1985), S. 427-433 
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial mutations ; Informational suppressors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ten nuclear suppressors (nam mutations) of the mitochondrial oxi1-V25 ochre mutation are characterized. They restore to some extent the functional form of cytochrome oxidase, as judged by the results of growth tests, cytochrome spectra, cytochrome oxidase activities, and electrophoresis of the products of mitochondrial translation. The nam mutants can suppress some mit − mutations mapping in four mitochondrial genes. They act on a number of chain-terminating mit − mutations. When grown on glycerol medium some double mutants nam x-V25 show an increased sensitivity to paromomycin, while the growth of others is stimulated by the drug. The nam mutants are probably omnipotent suppressors resulting from mutations in nuclear gene(s) specifying mitoribosomal protein(s).
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  • 48
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    Current genetics 9 (1985), S. 553-560 
    ISSN: 1432-0983
    Keywords: Yeast ; Ty element ; Recombination ; Gene conversion ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A haploid yeast mutant carrying a reciprocal translocation was analyzed. Cloning and comparison of sequences involved in the translocation event in wildtype and mutant revealed that the crossover between non-homologous chromosomes has occured within Ty sequences. By DNA sequence analysis it could be demonstrated that the reciprocal recombination event is accompanied by a short segment of non-reciprocal exchange (gene conversion) in the immediate vicinity of the crossover. Analysis of the translocation mutant and revertant isolates also indicated that the regulatory effect of Ty elements on adjacent genes can be modified by discrete changes within a Ty element.
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  • 49
    ISSN: 1432-0983
    Keywords: Mitochondria ; Mutation ; Yeast ; Selection ; Random drift
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Haploid yeast cells have about 50 copies of the mitochondrial genome, and a mutational event is unlikely to affect more than one of these at a time. This raises the question of how such cells, or their progeny, become fixed (homoplasmic) for the mutant alele. We have tested the roles of six hypothetical mechanisms in producing erythromycin-resistant mutant cells: (i) random partitioning of mitochondrial genomes at cell division; (ii) intracellular selection for mtDNA molecules of one genotype; (iii) intracellular random drift of mitochondrial allele frequencies; (iv) intercellular selection for cells of a particular mitochondrial genotype; (v) induction of mitochondrial gene mutations by the antibiotic used to select mutants; and (vi) reduction in the number of mitochondrial genomes per cell by the antibiotic. Our experiments indicate that intracellular selection plays the major role in producing erythromycin-resistant mutant cells in the presence of the antibiotic. In the absence of the antibiotic, the combined effects of random drift and random partitioning are most important in determining the fate of new mutations, most of which are lost rather than fixed. Our experiments provide no evidence for mutation induction or ploidy reduction by erythromycin.
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    Archives of microbiology 150 (1988), S. 309-312 
    ISSN: 1432-072X
    Keywords: Chloroquine ; Glycolytic enzymes ; Yeast ; Chloroquine and ATP/ADP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chloroquine at pH 8.0 and 10 mM concentration inhibits about 30% glucose consumption and ethanol formation in yeast cells. Out of the 11 glycolytic enzymes assayed, phosphoglycerate kinase and pyruvate decarboxylase have been found to be most sensitive to chloroquine. Next sensitive are hexokinase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase. Kinetic studies with the three kinases studied revealed competitive inhibition of chloroquine with ATP (hexokinase, phosphoglycerate kinase) or ADP (pyruvate kinase).
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    Archives of microbiology 151 (1988), S. 20-25 
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating ; Zygote formation ; Chloroquine ; Lysosomotropic agent ; Plasma membrane ; Cell fusion
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Haploid cells of opposite mating type of Saccharomyces cerevisiae conjugate to form zygote. During the conjugation process, the degradation or reorganization of the cell wall and the fusion of the two plasma membranes take place. Since chloroquine inhibits cellular events associated with the reorganization of the plasma membrane, the effect of the drug on conjugation was studied. Chloroquine at a concentration, at which cell growth was not retarded, inhibited zygote formation, while it did not affect other mating functions, such as sexual agglutination, production of and response to mating pheromone. Cells in a mating culture containing chloroquine formed no “prezygote” suggesting that they were not prepared for entering into fusion process. The inhibitory effect of chloroquine was reversible as cells formed zygote when they were washed after treatment with chloroquine. Zygote formation was unaffected in cells possessing chlorquine within vacuoles after incubation with the drug in complete medium (YPD) at pH 7.5, followed by washing. This suggests that chloroquine inhibits zygote formaton by adsorbing to the plasma membrane of S. cerevisiae.
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    Archives of microbiology 150 (1988), S. 37-41 
    ISSN: 1432-072X
    Keywords: Yeast ; Hexose transport ; Sugar ; Malate uptake ; 2,4-DNP ; Zygosaccharomyces bailii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When grown in fructose or glucose the cells of Zygosaccharomyces bailii were physiologically different. Only the glucose grown cells (glucose cells) possessed an additional transport system for glucose and malate. Experiments with transport mutants had lead to the assumption that malate and glucose were transported by one carrier, but further experiments proved the existence of two separate carrier systems. Glucose was taken up by carriers with high and low affinity. Malate was only transported by an uptake system and it was not liberated by starved malate-loaded cells, probably due to the low affinity of the intracellular anion to the carrier. The uptake of malate was inhibited by fructose, glucose, mannose, and 2-DOG but not by non metabolisable analogues of glucose. The interference of malate transport by glucose, mannose or 2-DOG was prevented by 2,4-dinitrophenol, probably by inhibiting the sugar phosphorylation by hexokinase. Preincubation of glucose-cells with metabolisable hexoses promoted the subsequent malate transport in a sugar free environment. Preincubation of glucose-cells with 2-DOG, but not with 2-DOG/2,4-DNP, decreased the subsequent malate transport. The existence of two separate transport systems for glucose and malate was demonstrated with specific inhibitors: malate transport was inhibited by sodium fluoride and glucose transport by uranylnitrate. A model has been discussed that might explain the interference of hexoses with malate uptake in Z. bailii.
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  • 53
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating reaction ; Zygote formation ; Mating pheromone ; Fatty acid ; Arachidonic acid
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    Notes: Abstract Effect of exogenous fatty acids on zygote formation in Saccharomyces cerevisiae was studied. Arachidonic and oleic acids considerably stimulated zygote formation, but other fatty acids tested, linoleic, linolenic, stearic and palmitic acids, did not. Pretreatment experiments with arachidonic acid showed that the stimulation of zygote formation by the fatty acid required the presence of mating pheromone.
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    Archives of microbiology 149 (1988), S. 261-267 
    ISSN: 1432-072X
    Keywords: Yeast ; Hanseniaspora uvarum ; Pichia kluyveri ; Killer toxin ; dsRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By heat treatment killer strains of the type K1 of Saccharomyces cerevisiae that are known to harbour dsRNA plasmids were completely cured, whereas only a small fraction of the clones of the killer type K2 had lost the dsRNA dependent killer character. The K2 killers but not the strains of killer type K1 were easily cured by cycloheximide. Killer strains of Hanseniaspora uvarum were not curable by heat treatment. Curing was successfull with cycloheximide or 5-fluorouracil. Two double-stranded RNA plasmids were detected in the killer strains of H. uvarum. The smaller dsRNA plasmid was absent in the strains that were cured of their killer character by 5-fluorouracil. The killer character of H. uvarum was transferred to S. cerevisiae by spheroplast fusion. The fusion products showing the killer character contained both dsRNA plasmids, obviously the smaller plasmid (M-dsRNA) carries the genes for killer toxin formation. Killer strains of Pichia kluyveri were not curable of their killer character, in these strains no dsRNA plasmids were detected.
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  • 55
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    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
    ISSN: 1476-5535
    Keywords: Osmotic pressure ; Intracellular ethanol ; Yeast ; Nutrient ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The intracellular accumulation of ethanol in yeast and its potential effects on growth and fermentation have been topics of controversy for the past several years. The determination of intracellular ethanol based on the exclusion of [14C]sorbitol to estimate aqueous cell volume was used to examine the question of intracellular ethanol accumulation. An intracellular accumulation of ethanol inSaccharomyces cerevisiae was observed during the early stages of fermentation. However, as fermentation continued, the intracellular and extracellular concentrations of ethanol became similar. Increasing the osmotic pressure of the medium with glucose or sorbitol was observed to cause an increase in the intracellular ethanol concentration. Associated with this was a decrease in yeast growth and fermentation rates. In addition, increasing the osmotic pressure of the medium was observed to cause an increase in glycerol production. Supplementation of the media with excess peptone, yeast extract, magnesium sulfate and potassium phosphate was found to relieve the detrimental effects of high osmotic pressure. Under these conditions, though, no effect on the intracellular and extracellular ethanol distribution was observed. These results indicate that nutrient limitation, and not necessarily intracellular ethanol accumulation, plays a key role during yeast fermentations in media of high osmolarity.
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  • 56
    ISSN: 1573-4927
    Keywords: wheat ; gliadins ; gel electrophoresis ; evolution ; genetic analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The gliadin composition of 78 spring durum wheat varieties has been studied by one-dimensional (Al-lactate,pH 3.1) and two-dimensional (first dimension, Al-lactate,pH 3.1; second dimension, sodium dodecyl sulfate-polyacrylamide gel) electrophoresis. Analysis of hybrids has shown that all components of the α zone of gliadin spectra are inherited together as blocks and are, probably, coded for by a cluster of tightly linked genes located on chromosome 6A. Fourteen variants of gliadin blocks have been identified, which can be classified into five families on the basis of component composition. All families but one have analogues among chromosome 6A-controlled blocks of bread wheat. The results indicate that some of the genome A diploid genotypes that were ancestors of durum wheats were also ancestors of bread wheats and that polyploid wheats were produced by repeated allopolyploidization events, as has been suggested earlier.
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  • 57
    ISSN: 1573-4927
    Keywords: wheat ; gliadins ; gel electrophoresis ; evolution ; genetic analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The gliadin composition of 78 spring durum wheat varieties has been studied by one-dimensional (Al-lactate,pH 3.1) and two-dimensional (first dimension, Al-lactate,pH 3.1; second dimension, sodium dodecyl sulfate-polyacrylamide gel) electrophoresis. Analysis of hybrids has shown that all components of the α zone of gliadin spectra are inherited together as blocks and are, probably, coded for by a cluster of tightly linked genes located on chromosome 6A. Fourteen variants of gliadin blocks have been identified, which can be classified into five families on the basis of component composition. All families but one have analogues among chromosome 6A-controlled blocks of bread wheat. The results indicate that some of the genome A diploid genotypes that were ancestors of durum wheats were also ancestors of bread wheats and that polyploid wheats were produced by repeated allopolyploidization events, as has been suggested earlier.
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  • 58
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    Plant molecular biology 10 (1988), S. 303-310 
    ISSN: 1573-5028
    Keywords: chloroplast gene ; monocot ; psbA ; transcription ; wheat ; 5′ end
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have mapped and cloned the wheat chloroplast gene (psbA) that encodes the 32 kd polypeptide of Photosystem II. The psbA gene is located in the large single copy region adjacent to one inverted repeat and is transcribed toward the latter. The sequence of the 5′ end of the wheat gene is homologous with dicot psbA genes. We have located the 5′ terminus of the wheat psbA RNA to a position 83 nt upstream of its coding region. The same psbA RNA species was capped in vitro by guanylyltransferase, establishing that its 5′ end is a transcription start site. Regions which resemble procaryotic -10 and -35 promoter elements are located immediately upstream of the wheat psbA transcription initiation site.
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  • 59
    ISSN: 1573-5028
    Keywords: wheat ; triplet protein (triticin) ; legumin ; amino acid sequence homology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have obtained several amino acid sequences from purified polypeptides of a wheat endosperm storage globulin previously described as ‘triplet’ protein. The amino acid sequence data supported by immunochemical analysis using anti-oat 12S globulin antibodies, provide definitive evidence that the triplet protein is homologous to pea legumin and related seed storage proteins of oats, rice and several dicotyledonous species. Thus, it is now proposed that the triplet protein of wheat be renamed ‘triticin’. The oat globulin antibodies also cross-reacted strongly with the high-molecular-weight (HMW) glutenin subunits which have been implicated in bread-making quality.
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  • 60
    ISSN: 1573-5028
    Keywords: mitochondrial DNA ; transfer RNA genes ; rearrangement ; recombination ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the course of isolating tRNA genes from wheat mtDNA, we have found the same tRNAPro gene in two different Hind III restriction fragments, H-P1 (0.7 kbp) and H-P2 (1.7 kbp). Sequences immediately flanking these duplicate genes are closely related, although not identical; sequence comparisons suggest that multiple rearrangements have occurred in the vicinity of the H-P2 tRNAPro gene, relative to the H-P1 version. The chimeric nature of H-P2 is emphasized by the presence of sequences that are also found upstream of the wheat mitochondrial 26S rRNA gene, as well as sequences derived from chloroplast DNA. Comparison of H-P2 with H-P1 plus upstream sequences provides some insight into possible molecular events that might have generated H-P2. In particular, such comparisons suggest a model in which the homologous sequences in H-P2 are seen to be derived from H-P1 plus upstream sequences as a result of an intragenomic, site-specific rearrangement event, followed by amplification of the product, its fixation in the mitochondrial genome, and subsequent sequence divergence (single base changes as well as insertions/deletions of up to 50 nucleotides). The results reported here implicate particular primary sequence motifs in certain of the rearrangements that characterize H-P2.
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  • 61
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    Plant molecular biology 10 (1988), S. 251-262 
    ISSN: 1573-5028
    Keywords: mitochondria ; mitochondrial DNA ; transfer RNA ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have begun a systematic search for potential tRNA genes in wheat mtDNA, and present here the sequences of regions of the wheat mitochondrial genome that encode genes for tRNAAsp (anticodon GUC), tRNAPro (UGG), tRNATyr (GUA), and two tRNAsSer (UGA and GCU). These genes are all solitary, not immediately adjacent to other tRNA or known protein coding genes. Each of the encoded tRNAs can assume a secondary structure that conforms to the standard cloverleaf model, and that displays none of the structural aberrations peculiar to some of the corresponding mitochondrial tRNAs from other eukaryotes. The wheat mitochondrial tRNA sequences are, on average, substantially more similar to their eubacterial and chloroplast counterparts than to their homologues in fungal and animal mitochondria. However, an analysis of regions ∼ 150 nucleotides upstream and ∼ 100 nucleotides downstream of the tRNA coding regions has revealed no obvious conserved sequences that resemble the promoter and terminator motifs that regulate the expression of eubacterial and some chloroplast tRNA genes. When restriction digests of wheat mtDNA are probed with 32P-labelled wheat mitochondrial tRNAs, 〈20 hybridizing bands are detected, whether enzymes with 4 bp or 6 bp recognition sites are used. This suggests that the wheat mitochondrial genome, despite its large size, may carry a relatively small number of tRNA genes.
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  • 62
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    Theoretical and applied genetics 76 (1988), S. 933-940 
    ISSN: 1432-2242
    Keywords: Gliadin ; Two-dimensional electrophoresis ; F1 monosomic analysis ; chromosomal assignment ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The gliadin components from four bread wheat cultivars: Chinese Spring, Capelle Desprez, Holdfast and Pane-247 and their monosomic F1s for the chromosomes of homoeologous groups 1 and 6 have been analyzed by two-dimensional (2-pH) polyacrylamide gel electrophoresis. Chromosomal location of gliadin genes and the allelic differences were well established by analyzing the different F1 monosomic hybrids, electrophoretical patterns and differences in relative staining intensity. A new gliadin encoded by a gene located on chromosome 6B in Chinese Spring is described. The two-dimensional patterns of gliadin in the other three varieties and the chromosomal location of their genes are reported for the first time. Relationships between gliadins in the two-dimensional patterns and the traditional system for their nomenclature are discussed.
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  • 63
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    Molecular genetics and genomics 214 (1988), S. 37-41 
    ISSN: 1617-4623
    Keywords: Homologous recombination ; UV induction ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have investigated the effects of UV irradiation of Saccharomyces cerevisiae in order to distinguish whether UV-induced recombination results from the induction of enzymes required for homologous recombination, of the production of substrate sites for recombination containing regions of DNA damage. We utilized split-dose experiments to investigate the induction of proteins required for survival, gene conversion, and mutation in a diploid strain of S. cerevisiae. We demonstrate that inducing doses of UV irradiation followed by a 6 h period of incubation render the cells resistant to challenge doses of UV irradiation. The effects of inducing and challenge doses of UV irradiation upon interchromosomal gene conversion and mutation are strictly additive. Using the yeast URA3 gene cloned in non-replicating single- and double-stranded plasmid vectors that integrate into chromosomal genes upon transformation, we show that UV irradiation of haploid yeast cells and homologous plasmid DNA sequences each stimulate homologous recombination approximately two-fold, and that these effects are additive. Non-specific DNA damage has little effect on the stimulation of, homologous recombination, as shown by studies in which UV-irradiated heterologous DNA was included in transformation/recombination experiments. We further demonstrate that the effect of competing single- and double-stranded heterologous DNA sequences differs in UV-irradiated and unirradiated cells, suggesting an induction of recombinational machinery in UV-irradiated S. cerevisiae cells.
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  • 64
    ISSN: 1617-4623
    Keywords: Tn5 ; Transposon excision ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have demonstrated that precise excision of bacterial transposon Tn5 can occur in the yeast, Saccharomyces cerevisiae. Tn5 insertions in the yeast gene LYS2 were generated by transposon mutagenesis made in Escherichia coli by means of a λ::Tn5 vector. Nine insertions of Tn5 into the structural part of the yeast LYS2 gene situated in a shuttle epsiomal plasmid were selected. All the plasmids with a Tn5 insertion were used to transform yeast strains carrying a deletion of the entire LYS2 gene or a deletion of the part of LYS2 overlapping the point of insertion. All insertions inactivated the LYS2 gene and were able to revert with low (about 10-8) frequencies to lysine prototrophy. Restriction analysis of revertant plasmids revealed them to be indistinguishable from the original plasmid without Tn5 insertion. DNA sequencing of the regions containing the points of insertions, made for two revertants, proved that Tn5 excision was completely precise.
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  • 65
    ISSN: 1617-4623
    Keywords: Aneuploidy ; Yeast ; Saccharomyces cerevisiae ; Methyl benzimidazol-2-yl carbamate ; Mitosis ; Meiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A system is described in which spontaneous and chemically-induced mitotic and meiotic hyperploidy can be assayed in the same diploid culture of Saccharomyces cerevisiae. Monitoring gene dosage changes at two loci on chromosome VIII, the test utilizes a leaky temperature-sensitive allele arg4-8 and low level copper resistance conferred by the single copy allele cup1 s. An extra chromosome VIII provides simultaneous increased dosage for both genes, resulting in colonies that are both prototrophic for arginine at 30° C and copper resistant. During mitotic cell divisions in diploids, spontaneous chromosome VIII hyperploids (trisomes and tetrasomes) occur at a frequency of 6.4×10-6 per viable cell. Among ascospores, the spontaneous chromosome VIII disome frequency is 5.5×10-6 per viable spore. The tubulin-binding reagent methyl benzimidazol-2-yl carbamate (MBC) elicits enhanced levels of mitotic and meiotic aneuploidy relative to control levels. The system represents a novel model for examining chromosome behavior during mitosis and meiosis and provides a sensitive and quantifiable procedure for examining chemically induced aneuploidy.
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  • 66
    ISSN: 1617-4623
    Keywords: Mutagen resistance ; Yeast ; Formaldehyde ; 4-Nitroquinoline-N-oxide ; Multi-copy plasmids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hyperresistance to 4-nitroquinoline-N-oxide (4-NQO) and formaldehyde (FA) of yeast strains transformed with the multi-copy plasmids pAR172 and pAR184, respectively, is due to the two genes, SNQ and SFA, which are present on these plasmids. Restriction analysis revealed the maximal size of SFA as 2.7 kb and of SNQ as 2.2 kb, including transcription control elements. The presence of the smallest 2.7 kb subclone carrying SFA increased hyperresistance to formaldehyde fivefold over that of the original pAR184 isolate. No such increase in hyperresistance to 4-NQO was seen with the smaller subclones of the pAR172 isolate. Disruption of the SFA gene led to a threefold increase in sensitivity to FA as compared with the wild type. Expression of gene SNQ introduced on a multi-copy vector into haploid yeast mutants rad2, rad3, and snm1 did not complement these mutations that block excision repair.
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  • 67
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    Molecular genetics and genomics 211 (1988), S. 41-48 
    ISSN: 1617-4623
    Keywords: RAD52 ; Repair ; Nuclease ; Antibody ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Extracts of Rad+ and radiation-sensitive (rad) mutants of the yeast Saccharomyces cerevisiae were examined for total Mg2+-dependent alkaline deoxyribonuclease activity and the presence of a nuclease that crossreacts immunologically with an antiserum raised against an endoexonuclease from Neurospora crassa, an enzyme exhibiting both deoxyribo- and ribonuclease activities. No significant differences were observed in total deoxyribonuclease activity between Rad+ and rad mutants. The antibody precipitable activity, however, was found to be 30%–40% of the total alkaline deoxyribonuclease activity in logarithmically growing Rad+ cells. Extracts of stationary phase cells were lacking in antibody precipitable activity. Using immunoblot methods, a 72 kDa crossreacting protein was identified from logarithmically growing cells that was absent from stationary phase cells. In all radiation-sensitive mutants examined, except rad52, at least 20% of total activity was precipitable. Extracts from logarithmically growing rad52 mutants, including a rad52::LEU2 insertion mutant, exhibited less than 10% of the Rad+ precipitable activity; however, some crossreacting material was detected. Although, the level of endo-exonuclease activity is influenced by the RAD52 gene, it is not the product of this gene. The total deoxyribonuclease and the antibody precipitable endo-exonuclease activities were also followed during meiosis. Unlike the Rad+ strain which had previously been shown to have increased levels of total and immunoprecipitable endo-exonuclease as cells underwent meiosis, the rad52 mutant exhibited no increases in either category of nuclease activity. Given the importance of the RAD52 gene in repair, recombination and mutagenesis, the endo-exonuclease may be a significant component of these processes.
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  • 68
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; tRNA synthetase gene ; Distribution of mutations ; Genetic drift
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present a statistical study of the nature and distribution of mutations along the NAM2 gene coding for the mitochondrial leucyl tRNA synthetase in Saccharomyces cerevisiae and S. douglasii (Herbert et al. 1988). Two important facts are observed: (1) the relative frequency of transitions and transversions is the same among silent substitutions and replacements. (2) The two kinds of mutations (silent substitutions and replacements) are distributed in the same way along the gene. This distribution is not random; the mutations are clustered and the clusters are regularly spaced along the gene. The NAM2 gene offers an example spaced along the gene. The NAM2 gene offers an example of recent divergence. We show that, in this case, the fixation of mutations is the result of genetic drift and of constraints on the nucleic acid sequence and not on that of the protein.
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  • 69
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; pre-mRNA splicing ; tRNA synthetase gene ; Incipient evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We studied the NAM2 genes of Saccharomyces douglasii and Saccharomyces cerevisiae, and showed that they are interchangeable for all the known functions of these genes, both mitochondrial protein synthesis and mitochondrial mRNA splicing. This confirms the prediction that the S. douglasii NAM2D gene encodes the mitochondrial leucyl tRNA synthetase (EC 6.1.1.4). The observation that these enzymes are interchangeable for their mRNA splicing functions, even though there are significant differences in the intron/exon structure of their mitochondrial genome, suggests that they may have a general role in yeast mitochondrial RNA splicing. A short open reading frame (ORF) precedes the synthetase-encoding ORF, and we showed that at least in S. cerevisiae this is not essential for the expression of the gene; however, it may be involved in a more subtle type of regulation. Sequence comparisons of S. douglasii and S. cerevisiae revealed a particularly interesting situation from the evolutionary point of view. It appears that the two yeasts have diverged relatively recently: there is remarkable nucleotide sequence conservation, with no deletions or insertions, but numerous (albeit non-saturating) silent substitutions resulting from transitions. This applies not only to the NAM2 coding regions, but also to two other ORFs flanking the NAM2 ORF. The regions between the ORFs (believed to be intergenic regions) are much less conserved, with several deletions and insertions. Thus S. douglasii and S. cerevisiae provide an ideal system for the study of molecular evolution, being two yeasts “caught in the act” of speciation.
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  • 70
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    Molecular genetics and genomics 213 (1988), S. 400-408 
    ISSN: 1617-4623
    Keywords: Yeast ; DNA repair ; RAD3 gene expression ; Fusion proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cloned RAD3 gene of Saccharomyces cerevisiae was tailored into expression vectors for overexpression of Rad3 protein in Escherichia coli and in yeast. In both organisms the overexpressed protein is detected as a species of molecular weight ca. 90 kDa, the size expected from the sequence of the cloned gene. The protein overexpressed in E. coli is largely insoluble; however the insoluble fraction was used to generate affinity-purified polyclonal antisera which proved to be powerful reagents for the initial characterization of Rad3 protein expressed in yeast. These studies showed that: (1) when overexpressed in yeast most of the Rad3 protein is detected in the soluble fraction of cell extracts; (2) endogenous Rad3 protein is untransformed cells is also ca. 90 kDa in size and is located in the cell nucleus; (3) Rad3/β-galactosidase fusion protein partially purified on an affinity matrix is associated with DNA-dependent ATPase activity that is inhibited in the presence of anti-Rad3 antibodies, suggesting that Rad3 protein is an ATPase; and (4) Rad3 antibodies cross-react with two electrophoretically distinguishable polypeptides present in the nuclear fraction of human cells, and with a single polypeptide in extracts of Drosophila cell.
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  • 71
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed-phase ; Codeine phosphate drug substance ; Opium alkaloids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A rapid procedure for the evaluation of codeine phosphate drug substance derived from poppy straw or opium concentrate is described. The gradient HPLC procedure employs a pH 2.5 phosphate buffer, methanol and acetonitrile mobile phase at 40°C and a μBondapak C-18 column. Eight opium alkaloids, including the four major alkaloids, are separated from each other. The paper describes details of the assay procedure and presents data documenting the linearity, specificity, precision, and sensitivity of the method.
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  • 72
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; External variance determinations ; Column and system compatibility ; Microbore and fast LC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The demand for faster, more economic and convenient high-efficiency LC analyses has led to the development of microbore and fast columns. However, to utilize their benefits, the low column dispersions produced by these new technologies necessitate low external variances in the total system. This paper describes the determination of the external variance of microbore, fast and conventional LC systems by a column method. Compatibility with the relevant column technologies is evaluated by a method involving the comparison of the external and column dispersions. Direct comparison of chromatograms of an 8-component mixture, on a set of columns of various diameters and lengths, shows the relative speed, economy and sensitivity obtainable in practice. The problem of reduced sample loadability is considered and the practical solution of peak compression illustrated.
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  • 73
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Retention prediction system ; UV multichannel detector ; Polycyclic aromatic hydrocarbons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The retention prediction system assisted by a micro-computer coupled with an UV multichannel detector has been investigated in the separation of polycyclic aromatic hydrocarbons. The method offered very precise identification of components in NBS-SRM-1647 sample without any standard reference substances. It has been proven that the system makes very precise, non-pollutive and convenient environmental analysis possible.
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  • 74
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    Chromatographia 20 (1985), S. 313-322 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Tetracyclines antibiotics ; Quantitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The use of high-performance liquid chromatography (HPLC) for the identification and determination of tetracycline antibiotics is reviewed. HPLC chromatograms provide fast identification by retention time, tR, and precise quantitation by measurement of peak height or peak area. For separation of tetracycline compounds, most HPLC methods use reversed-phase C18 or C8 columns and UV detection. The HPLC solvent system should have a pH of about 6 to prevent steric changes in the tetracycline molecule. For accurate quantitation it is necessary to avoid tailing and this is accomplished by adding a zwitter ion to the solvent system. Methanol and acetonitrile are frequently used as organic modifiers in these solvent systems. In a single analysis, HPLC methods can be used to separate as many as nine or ten commercially used tetracycline compounds and to determine four to five tetracyclines in commercial tetracycline preparations or in biological fluids.
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  • 75
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Resolution of optical isomers ; Chiral column ; Pyrethroids ; Fenvalerate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The separation of the optical isomers of fenvalerate [cyano(3-phenoxyphenyl)methyl 2-(4-chlorophenyl)-3-methylbutyrate] has been carried out by high-performance liquid chromatography (HPLC) on a chiral column with (R)-N-3,5-dinitrobenzoyl-phenyl-glycine (DNBPG) covalently bonded on aminopropyl silica and eluted with mixtures of methanol, 2-propanol, and hexane. The system was applied to the analysis of Pydrin®, an emulsifiable concentrate formulation of fenvalerate, and to the residue analysis of fenvalerate in milk samples. For the analysis of Pydrin® the only requirement was the proper dilution of Pydrin® with hexane. For the analysis of residues in milk, fenvalerate was extracted with hexane after precipitating the milk proteins with acetonitrile and removing the precipitate by filtration; the hexane extract was concentrated to small volume and filtered before being analyzed by HPLC.
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  • 76
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    Chromatographia 20 (1985), S. 487-494 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Size-exclusion chromatography ; Controlled-porosity glasses ; Boron-enriched glass surface ; Adsorptive properties
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Controlled-porosity glasses (CPG) are sieves for macromoleculars, very widely applied in chromatographic columns for the separation of polymers and biopolymers by means of size-exclusion chromatography (SEC) and affinity chromatography. This paper deals with the influence of the thermal treatment of CPG on the elution of polymers in SEC columns. The problem is examined for a few mobile phases and for glasses having different porosities. Additionally, the SEC results obtained are compared with the adsorption properties of the glases investigated.
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  • 77
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    Chromatographia 25 (1988), S. 31-36 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Radioactivity detector ; Solid scintillator ; Fibre-packed flow cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new type of flow-cell radiation detector for use in liquid chromatography which is packed with aligned scintillator fibers is described. A primary advantage of the fiber packed cell is that light generated by the scintillator is absorbed to a much less extent by the fibers than by the powder scintillator used in conventional flow cells. A detection efficiency of 55% has been obtained for carbon-14 using 0.1-mm diameter hand-pulled glass fibers. Computer modeling has shown that even better results can be obtained by using smaller and more uniform diameter fibers which will allow better packing. The fiber cell also demonstrates back pressures which are a factor of 50 less than the conventional cell and much less susceptibility to absorption of compounds because of its lower surface area.
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  • 78
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    Chromatographia 25 (1988), S. 854-860 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polystyrene ; Gradient elution ; Turbidimetry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polystyrene samples of narrow molecular-weight distribution have been eluted according to their molecular weight from columns packed with bare silica Si50, phenyl, or C18 bonded phase by gradients of methanol and tetrahydrofuran (THF) or ofiso-octane and THF. Among the six combinations investigated,iso-octane/THF with a silica column formed a proper normal-phase system whereas methanol/THF with a C18 column formed a proper reversed-phase system. The combinations of C18 column andiso-octane/THF or of Si50 column and methanol/THF gradient did not correspond to the approved polarity rules in high-performance liquid chromatography but were nevertheless effective in separating polystyrene mixtures by molecular weight. Methanol andiso-octane are nonsolvents for polystyrene whereas THF is a solvent. The solubility of polystyrene as a function of molecular weight and concentration was determined by means of turbidimetric titration of solutions in THF with the nonsolvents used in the gradients. The solubility and elution characteristics were almost identical on C18 columns or in methanol/THF combinations. The elution from phenyl bonded phase and Si50 columns usingiso-octane/THF gradients required more THF than the solubility experiments. Information is also given on the occurrence of multimodal elution patterns.
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  • 79
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polycyclic aromatic compounds ; Alkylated PAC ; Retention characteristics ; Normal phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Retention characteristics of series of polymethyl and mono-n-alkyl derivatives of benzene and pyrene, and also of parent polycyclic aromatic hydrocarbons (PAH), were studied using silica and aminopropyl- and cyanopropyl-modified silica. Differences in the selectivities for the studied compound groups were found between the three phases. Deviations from linear behaviour in plots of log (k′)vs. carbon number were observed for the methyl series. These are explained in terms of differences in π-electron delocalisation within the aromatic ring systems. Further, the effect of methyl substitution on selectivity decreased with an increasing number of aromatic rings. Results were obtained which indicated that the primary adsorption site in a cyano column used in normal phase mode, at least for PAH molecules, is the cyano group.
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  • 80
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    Chromatographia 25 (1988), S. 43-50 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phase ; Void volume ; Adsorption isotherms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In reversed-phase liquid chromatography with n-alkyl bonded silica, the dead volume (V0) of the column is theoretically indeterminate owing to adsorption of organic modifier on n-alkyl chains and of water on silanol groups. With binary mobile phases, retention volumes of the mobile phase components and of their deuterated species are relaeed to the adsorption isotherms and V0 by equations which can be solved with some assumptions on the adsorbed layer composition. Methanol-water and acetonitrile-water systems are studied. As the experimental excess isotherm shows a linear part in the concentration range 50–80% in organic modifier, the hypothesis of an adsorbed layer of constant composition in this range is possible. When increasing the water content of the mobile phase, adsorption of water occurs up to saturation of silanol groups. Then the assumption of a constant water content for a mobile phase having more than 50% of water is applied. With the hypothesis of a constant adsorbed content of organic modifier when the eluent has more than 80% of organic modifier, V0 and the absolute isotherms are calculated over the entire range of mobile phase composition. Experimental retention behavior of the mobile phase components are totally explained by these V0 determinations. The retention times of commonly used V0 markers are compared with V0 values. It is shown that, when buffering the eluent, no visible effect on the distribution equilibrium is observed, so that injection of concentrated potassium nitrate is a convenient method to measure V0. With a few solutes with are UV detectable it is possible to measure V0 whatever the mobile phase composition in methanol-water and acetonitrile-water systems.
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  • 81
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Stationary phase solubility parameters ; Cyano-silica ; Amino-silica ; Diol-silica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Extended solubility parameters have been determined for aminopropyl, cyanopropyl and 1,2-dihydroxypropyl propyl ether (diol) normal bonded-phase HPLC columns. Parameters were calculated from both the retention data of solutes (partition model) and empirically determined solvent strengths (adsorption-displacement model). The use of solvent strengths to calculate solubility parameters for these silica-based bonded stationary phases appears to be superior, since this technique avoids many of the problems that arise from their inherent heterogeneity. Normalized solubility parameters were also used to position these columns on a stationary phase selectivity triangle. The amino and cyano phases appear in regions of the tringle expected based on the properties of the pure liquids, but an ether linkage in the diol apparently neutralizes some of the acidity expected from this phase.
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  • 82
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Macrolide antibiotic azithromycin ; Intermediates of azithromycin ; Optimization of separation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary HPLC is the most convenient method for the assay of the azithromycin (10-dihydro-10-deoxo-11-methyl-11-azaerythromycin A), a semisynthetic macrolide antibiotic and its reaction intermediates. Data concerning the effects of pH, temperature and column type are presented. The analytical procedures enabling the reliable assay of azithromycin and its intermediates, as well as other impurities are defined. Use of a reversed-phase octadecyl column, a pH of 9.3–9.5, and isocratic mode at ambient temperature are the best conditions of analysis.
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  • 83
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Metabolic disorders ; Xanthine and 2,8-dihydroxyadenine ; Urine analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary 2,8-Dihydroxyadeninuria and xanthinuria are inborn enzyme disorders which must be under lifelong therapeutic control. Quantitative determination of 2,8-DHA and xanthine using reversed phase HPLC was performed. A new application for the separation of purines shortens analysis-time. Examples of the determination of 2,8-DHA and xanthine before and under certain therapies are shown. Long term monitoring of the patients offers the possibility of a reliable prophylaxis against stone recurrence.
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  • 84
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Indirect detection of non-electrolytes ; Simulation of induced peaks ; RP systems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Induced peaks observed in indirect detection of nonelectrolytes in high-performance liquid chromatography are simulated. The equation representing the signal intensity of the induced peaks is derived, and it is verified by the experimental results. The peak area is proportional to (K b ′ +1) k a ′ /| a ′ —k b ′ |, where k a ′ ad k b ′ are the capacity factors of the visualization agent and the analyte, respectively.
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  • 85
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    Chromatographia 25 (1988), S. 162-166 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion chromatography ; Analysis of metal ions ; Post column derivatisation ; Amperometric detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for determination of metal ions subsequent to separation by ion chromatography has previously been developed. The method is based on Indirect Amperometric Detection whereby the decrease in the oxidation current, due to a dithiocarbamate ligand added post column, is monitored. Upon elution from the chromatographic column the metal ions are complexed by the ligand. As the complexes formed are electroinactive at the applied potential, the background current decreases according to the metal ion concentration. The method developed in this work involves addition to the reagent of Zn(II) as a auxiliary metal ion to displace the analyte metals from the chromatographic eluent ligand complexes after separation. Sodium bis(2-hydroxy-ethyl) dithiocarbamate was used as the postcolumn derivatising reagent. The addition of Zn(II) to the reagent causes some unforeseen behaviour in the chromatographic system.
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  • 86
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    Chromatographia 25 (1988), S. 199-204 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Column switching ; Catecholamines ; Urine ; Serum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary We developed an automated, two-column HPLC-method that can be used routinely to quantify the catecholamines norepinephrine and epinephrine in body fluids. The method is based on a new, laboratory-prepared SEC-HPAC column material, on the application of a microprocessor-controlled column-switching technique, and on the use of an integrated reaction system for postcolumn derivatization and trihydroxyindol-fluorescence monitoring. It allows the direct injection (upto 500 μl) of an appropriate biological fluid and distinguishes on its integrated sample-processing mode, its practicability, its chemoselectivity and-specivity, its detection limit (2 pg), its within- and between-run precision and its speed of analysis.
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  • 87
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Pheophytinatonicke(II) ; Chlorophyll ; Pheophytin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary HPLC determination of pheophytinatonicke(II) (Pheo-Ni) prepared by the replacement of magnesium(II) in chlorophyll with nicke(II) is described. The good separation of PheoNi was obtained by using chemically bonded C18 as the stationary phase and acetone-methanol (50∶50, vol/vol) as the mobile phase. Conventional spectrophotometric method was also used for the determination of PheoNi. For the synthetic samples prepared by mixing (pheophytinato a) nicke(II) [(Pheo-a) Ni] and (pheophytinato b) nicke(II) [(Pheo-b) Ni], analytical values obtained by the spectrophotometric method were very high compared to those obtained by HPLC. In the proposed HPLC method, (Pheo-a) Ni and (Pheo-b). Ni could be determined in the concentration range of 0.028–30μg/ml and 0.038–30μg/ml with relative standard deviations (n=10) of 3.1% and 0.8%, respectively.
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  • 88
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Flavonoid compounds ; Structure-retention relationships ; Reversed-phase systems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Reversed-phase HPLC retention behaviour of different flavonoid compounds in different columns and with a range of methanol-buffer eluents was examined. Repeatibility and reproducibility of logk′ and Δlogk′ values were analyzed. Sharp group contributions, expressed as Δlogk′, were observed. These did not depend significantly on acid modifier type, column packing characteristics (C8 and C18) and dimensions (5,7 and 10μm) or the percentage of methanol in the mobile phase. A list of twelve group contributions is presented which is in agreement with literature data.
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  • 89
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    Chromatographia 25 (1988), S. 443-446 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Large scale separations ; Dissolution of silica ; Metallic contaminants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Possible contamination of purified product by the silica matrix is of considerable importance when considering process scale high-performance liquid chromatography (HPLC) as a separation methodology. The stability of silica supports in normal phase, reversed phase and affinity chromatography modes has been examined with organic solvents and water. Both inorganic and organic components were found, contributed by the support, and the chromatographic and distillation equipment. In all cases, based on assumed values of production, they were found to contribute less than 0.01% of impurities. In critical cases crystallisation or dialysis may be considered as a last stage of purification, with re-chromatography to maximise yields.
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  • 90
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fast atom bombardment mass spectrometry ; Nonionic detergents
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Microcolumn liquid chromatography (LC) combined with fast-atom-bombardment mass spectrometry (FABMS) was applied to the analysis of nonionic detergents. The porous silver filter attached to the top of the capillary fused-silica tubing worked as the interface for coupling of micro LC and FABMS.
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  • 91
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    Chromatographia 25 (1988), S. 577-581 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Process scale ; Examples of separations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The flexibility of process scale high performance liquid chromatography is demonstrated by three examples of reversed-phase separations (a) a standard purification (b) isolation of trace compounds, and (c) recovery from crystallisation mother liquors. It was shown that a material, thought to be acceptably pure, contained a toxic impurity, and a remaining impurity, a previously unseen component of potential therapeutic interest. Favourable cost data were indicated for example (3).
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  • 92
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    Chromatographia 25 (1988), S. 717-720 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed-phase chromtography ; Antifungla agent ; Pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liqid chromatographic method is reported for the measurement of miconazole in systemic, fungal infectious patients. Pharmacokinetic data are presented for a single patient receiving miconazole therapy. Sample preparation involves protein precipitation by acetonitrile (1:1, vol/vol). Analyses are carried out on a reversed-phae chromatographic system using octadecylsilane stationary phase: a mobile phase consisting of 0.05 M acetate buffer (pH 7.4)acetonitrile (20:80, vol/vol) is used to elaute miconazole is quantified on the basis of ultraviolet absorption at 220 nm. The precision of the method ranged from 3.21% at 0.5 mg/L to 0.85% at 2.0 mg/L. The limit of quantification was established as 0.1 mg/L. Interference from other drugs that are co-adimistered such as amphotericin B, 5-fluorocytosine of ketoconazole and most other comonly encountered drugs was not observed.
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  • 93
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fractional factorial experimental designs ; Ruggedness test ; Method validation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Recently much attention has been directed to the validation of analytical methods. The level of validation will depend on the method application. If a method is developed for general use, for instance by different analysts, instruments or laboratories, then the effect of these changes need evaluation. This is normally referred to as ruggedness testing, where the limitations of the method to changes in the specified conditions are examined. This paper examines the use of Plackett-Burman fractional factorial experiment designs for ruggedness testing in high performance liquid chromatography (HPLC). The ability of these designs to identify confounding effects is applied to the stability indicating analysis of Salbutamol and its major degradation product, AH4045. The results show that confounding effects, proving extremely pertinent to HPLC, can be identified. The assay proved rugged to most tested changes although not to the use of column packing material from different manufacturers.
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  • 94
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Alkyl-bonded phases ; Retention behaviour ; Ligand density ; n-alkyl chain length
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two series of bonded phases were synthesized employing LiChrospher Si 100, 10 μm and n-alkyldimethylmonochlorosilanes as silanizing reagents. In series A the n-alkyl chain length, n, of the bonded phase was varied between 1 and 20 at a constant ligand density of 3.5±0.2μmol·m−2. In series B the ligand density, d, was gradually changed from 0 to 4.1μmol·m−2 on the C1, C4, C6, C8 and C18 bonded phases, respectively. The capacity factors of benzoic acid esters and anilines as solutes were found to increase linearly with the n-alkyl chain length of packings at constant eluent composition (series A) up to a so-called critical chain length, ncrit, where the capacity factor remained constant. ncrit was in the range from 11 to 14. The same pattern was observed when plotting the solute capacity factor against the ligand density of the packing at constant n and constant eluent composition (series B). The critical ligand density, αcrit, varied between 2.3 and 3.2 μmol·m−2 depending on n and the solute. Furthermore, solute retention was slightly higher on RP packings with an even number of carbon atoms in the alkyl chain than on those with an odd number. These findings are consistent with the results of Berendsen and de Galan (J. Chromatogr., 196, 21 (1980)), Dill (J. Phys. Chem., 91, 1980 (1987)) and Simpson and Lau (to be published). The observed phenomena reflect the dynamic structure of RP silicas which are related to the mobility of solvated n-alkyl chains. Due to the lack of a precise conformational analysis of the surface of RP silicas, a semiquantitative model was applied to interprete the described dependencies.
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  • 95
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Yeast ; Copy number ; Thymidine kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 2 μm circle-based chimaeric plasmid containing the yeast LEU2 and the Herpes Simplex Virus type 1 thymidine kinase (HSV-1 TK) genes was constructed. Transformants grown under selective conditions for the LEU2 gene harboured the plasmid at about 15 copies per cell whilst selection for the HSV-1 TK gene led to an increase to about 100 copies per cell. Furthermore, the plasmid copy number could be controlled by the stringency of selection for the TK gene, and the increase in TK gene dosage was reflected in an increase in intracellular thymidine kinase activity. The mitotic stability of the plasmid in “high-copy” and “low-copy” number cells was determined. “High-copy” number cells showed a greater mitotic stability. The relationship of TK expression to plasmid copy number may be useful for the isolation of plasmid copy number mutants in yeast and the control of heterologous gene expression.
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  • 96
    ISSN: 1617-4623
    Keywords: Yeast ; Ty elements ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We examined Ty-mediated genomic rearrangements in three related mitotically dividing haploid yeast strains having the same configuration of Ty elements in the CYC1-sup4 interval of chromosome X. Surprisingly, quite different types and frequencies of rearrangements were found in the three strains. In one strain we found only Ty-mediated deletions, which occurred with a frequency of about 1×10-6. Another strain yielded similar deletions, but approximately one-third of these were accompanied by adjacent Ty-mediated inversions. A third strain was found to have an extremely high rate of inversion/reinversion between two of the three Ty elements. This rate was conservatively estimated to be 1.4±0.2×10-2 per cell per generation, which is at least 2 orders of magnitude higher than previously reported values for Ty-mediated rearrangements. These data provide evidence that local regions of the genome can, in some cases, be much more fluid than had been previously believed.
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  • 97
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    Molecular genetics and genomics 211 (1988), S. 492-498 
    ISSN: 1617-4623
    Keywords: Cytochrome c oxidase ; Mitochondria ; PET genes ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified and isolated a novel yeast nuclear gene (SCO1) which is essential for accumulation of the mitochondrially synthesized subunit II of cytochrome c oxidase (CoxII). Analysis of the mitochondrial translation products in a sco1-1 mutant reveals a strong reduction in CoxII. Examination of mitochondrial transcripts by Northern blot hybridization shows that transcription and transcript maturation of OXI1, the gene coding for CoxII, is not affected. Therefore the SCO1 gene product must be involved in a post-transcriptional step in the synthesis of CoxII. We have isolated a 1.7 kb DNA fragment from a yeast gene bank which carries the functional SCO1 gene. Two RNA species of 0.9 kb and 1.2 kb, respectively, hybridize with this DNA fragment, which is localized on chromosome II. Cells whose chromosomal 1.7 kb fragment has been replaced by the yeast URA3 gene fail to accumulate CoxII and in addition subunit I of cytochrome c oxidase (CoxI). The possibility that the SCO1 gene product is bifunctional, i.e. required for both CoxI and CoxII accumulation, is discussed.
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  • 98
    ISSN: 1617-4623
    Keywords: Yeast ; DNA primase-polymerase complex ; Temperature sensitive mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cloned DNA polymerase I gene has been used to map the POL1 locus on the left arm of chromosome XIV, between MET4 and TOP2. Temperature-sensitive mutants in POL1 have been obtained by in vitro mutagenesis of the cloned gene and in vivo replacement of the wild-type allele with the mutated copy. Physiological and biochemical characterization of one temperature-sensitive mutant (pol1-1) shows that cells shifted to the non-permissive temperature can complete one round of cell division and DNA replication before they arrest. Analysis of DNA polymerase I in crude extracts and in partially purified preparations indicates that the pol1-1 mutation results in a conformational change and affects the stability of the DNA primase-polymerase complex.
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  • 99
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    Molecular genetics and genomics 215 (1988), S. 46-52 
    ISSN: 1617-4623
    Keywords: Yeast ; pGKL plasmids ; Palindrome-hairpin linear plasmids ; the ORF1 gene ; DNA polymerase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast Kluyveromyces lactis haboring linear DNA plasmids pGKL1 and pGKL2 exhibits killer and killer-resistant phenotypes. Two new linear plasmids pK192L and pK192S were found in the weak killer mutant KUV192 induced by UV irradiation. pK192S was always accompanied by pK192L in subclones of KUV192. Both plasmids were derived from pGKL1 by deletion of the large right part of it. pK192L was 4.9 kb in size and had a palindromic structure consisting of 2.35 kb inverted terminal repetitions and a 215 base unique sequence. Analysis of denatured and renatured DNA strands suggested that pK192S was a hairpin-like form of pK192L. The pK192 plasmids were maintained only in cells haboring either pGKL1 or pGKL1S in addition to pGKL2 and competed with pGKL1 or pGKL1S for their maintenance. Since no complete ORF1 was conserved in pK192 plasmids, these results lead to the conclusion that the ORF1 gene is necessary for the replication and/or maintenance of pGKL1.
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  • 100
    ISSN: 1617-4623
    Keywords: Phosphoglucose isomerease ; Regulation ; Metabolism ; Glycolysis ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned and sequenced the PGI1 gene, encoding phosphoglucose isomerase (E.C.5.3.1.9), from Saccharomyces cerevisiae. The nucleotide sequence predicts subunits of 554 amino acids with a molecular weight of 61230. Both the size and amino acid composition correlate well with measurements from purified protein. We have compared the PGI1 protein with the predicted sequence for pig muscle PGI. In spite of some evolutionary divergence the proteins are very similar and there are some highly conserved regions, two of which have been implicated in the active site. It has been suggested that PGI exists in two or more isozyme forms in S. cerevisiae and analogy with ADR2/ADC1 suggests that such PGI isozymes might also be differentially regulated during glycolytic/gluconeogenic growth. We have used accurate quantitation of PGI1 mRNA and gene fusions of PGI1 to the lacZ gene of Escherichia coli to show that PGI1 transcription is regulated neither between glycolytic and gluconeogenic growth nor between exponential and stationary phase. The complete lack of PGI activity in PGI1 deletion mutants and of differential regulation suggests that the isozymes of PGI might result merely from processing of the PGI1 gene product.
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