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  • 1
    ISSN: 1432-0983
    Keywords: Key words  Agaricus bisporus ; Homologous recombination ; Exo-β-1 ; 3-glucanase ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   Homologous integration was studied in the common mushroom, Agaricus bisporus, using a plasmid (pHAG3-1) carrying the hygromycin-resistance gene and a 3.2-kb genomic fragment from A. bisporus. Homologous integration was found in 30–60% of the transformants obtained with pHAG3-1 linearized at three different positions within the homologous sequence, generating either blunt, 5′- or 3′-protruding ends. The genomic fragment was found to contain two homologous open reading frames in tandem, which showed 60% similarity to exo-β-1,3-glucanases from Saccharomyces cerevisiae and Candida albicans. The level of the corresponding mRNA is low in the vegetative mycelium and relatively high in fruiting bodies. In the vegetative mycelium of a transformant with tandemly integrated pHAG3-1 plasmids at the homologous position, exoglucanase mRNA was strongly increased without any apparent effect on growth rate or morphology.
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  • 2
    ISSN: 1365-2494
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Plants from a Festuca pratensis variety which had caused‘eczema solare’among lambs in The Netherlands revealed the presence of endophytic Acremonium after microscopic evaluation. Combined gas chromatographic-mass spectrometric analysis of extracts from seeds of these plants showed the occurrence of N-formyl- as well as N-acetyl-loline which are formed as a result of the plant-fungus association.Endophyte-infected Lolium perenne plants produced lolines only at elevated temperatures while non-infected plants failed to synthesize these alkaloids under identical environmental conditions. Results indicate that presence of endophytic fungi together with growth at temperatures above 23°C is a prerequisite for induction of alkaloids detected in perennial ryegrass
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  • 3
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transformed hairy root cultures of Beta vulgaris L., grown in petri-dishes, were inoculated with a suspension of surface-sterilized larvae of the beet cyst nematode Heterodera schachtii Schm. Larvae developed into both male and female adults. Juvenile larvae were hatched from the newly-formed cysts, indicating that fertilization had occurred. Results of a glasshouse test showed that H. schachtii did not lose its pathogenicity after being cultured on transformed roots. This technique can be developed further for the mass-propagation of sedentary nematodes and for the in vitro storage of isclates.
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  • 4
    ISSN: 1432-2048
    Keywords: Alginate entrapped cells ; l-Dihydroxyphenylalanine ; Mucuna ; l-Tyrosine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The optimization of the biotransformation of l-tyrosine into l-dihydroxyphenylalanine (DOPA), and of formyl-tyrosine into formyl-DOPA by alginate-entrapped cells of Mucuna pruriens is reported. This optimization is discussed in terms of parameters that are relevant for the entrapped cell system (charge of the beads with cells, bead diameter, permeation of the cells) and some parameters that are relevant for the enzymatic transformation (pH, pO2, concentration of ascorbate). The optimization experiments resulted in the description of a biotransformation system which operates at a constant redox potential. With this transformation system a transformation efficiency of 70% could be obtained, at a biotransformation-rate of 435 μmol·h-1·g-1 (DW of cells) at a substrate concentration of 19 mM.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Planta 165 (1985), S. 264-268 
    ISSN: 1432-2048
    Keywords: Amino acid (biotransformation) ; D, L-Ethionine ; L-Dihydroxyphenylalanine ; Mucuna ; p-Fluorophenylalanine ; Phenoloxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of limitating nitrogen-containing compounds in the medium and of adding the amino-acid analogues p-fluorophenylalanine and ethionine on both phenoloxidase activity and the accumulation of L-3,4-dihydroxyphenylalanine (L-DOPA) are reported for cell suspension cultures of Mucuna pruriens. Nitrogen limitation of the cultures, or the addition of p-fluorophenylalanine or ethionine to the culture medium resulted in an increased phenoloxidase activity. There appeared to be an inverse relationship between phenoloxidase activity and the acccumulation of L-tyrosine into L-DOPA by alginate-entrapped cells occurred at a higher rate when phenoloxidase activity was increased.
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  • 6
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known α-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable α-amylase. The half-time of inactivation of this α-amylase was 5.1 h at 80°C and 2.4 h at 90°C. The temperature optimum for activity of the α-amylase was 70°C; the pH optimum for activity was relatively low, in the range 5.5–6.0. α-Amylase synthesis was regulated by induction and repression mechanisms. An inverse relationship was found between growth rate and α-amylase production. Low starch concentrations and low growth temperatures were favourable for enzyme production by the organism. At the optimal temperature for growth, 65°C, the α-amylase was a growth-associated enzyme. The optimal temperature for α-amylase production, however, was 40°C, with α-amylase increasing from 3.9 units (U)/ml to 143 U/ml when lowering the growth temperature from 65°C to 40°C. Maximal α-amylase production in a batch fermentor run at 65°C was 102 U/ml, which was 26-fold higher than in erlenmeyer flasks at 65°C. The dissolved O2 concentration was found to be a critical factor in production of the α-amylase.
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  • 7
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known α-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable α-amylase. The half-time of inactivation of this α-amylase was 5.1 h at 80°C and 2.4 h at 90°C. The temperature optimum for activity of the α-amylase was 70°C; the pH optimum for activity was relatively low, in the range 5.5–6.0. α-Amylase synthesis was regulated by induction and repression mechanisms. An inverse relationship was found between growth rate and α-amylase production. Low starch concentrations and low growth temperatures were favourable for enzyme production by the organism. At the optimal temperature for growth, 65°C, the α-amylase was a growth-associated enzyme. The optimal temperature for α-amylase production, however, was 40°C, with α-amylase increasing from 3.9 units (U)/ml to 143 U/ml when lowering the growth temperature from 65°C to 40°C. Maximal α-amylase production in a batch fermentor run at 65°C was 102 U/ml, which was 26-fold higher than in erlenmeyer flasks at 65°C. The dissolved O2 concentration was found to be a critical factor in production of the α-amylase.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; Beta vulgaris ; sugar beet ; regeneration ; shooter mutants ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Beta vulgaris plants were found to be susceptible to Agrobacterium tumefaciens strains carrying octopine Ti-plasmids after wounding of GA3 elongated stems or of hypocotyls. Tumors could be isolated and cultured aseptically. The tumor marker, octopine synthase (Ocs) activity, was present demonstrating the applicability of the Agrobacterium system for transfer of genetic information. For the production of transgenic plants two procedures were tested: inoculation of explants derived from cotyledons and hypocotyls of two weeks old seedlings and a leaf-disc procedure. The first method yielded both octopine positive calli as well as shoot regeneration on the six genotypes tested. In most cases, regeneration occurred from pre-existing, non-transformed meristems. The presence of Ocs activity could not be demonstrated in these shoots, although in one case octopine positive callus was formed at the base of the shoot, suggesting a chimeric structure of the plantlet or T-DNA genes, which were silent within the shoot and became active again in proliferating callus. The leaf-disc method did not give rise to direct or indirect regeneration, but transformed callus proliferated on the leaf edges. Optimal transformation frequencies were dependent on B. vulgaris genotype and Agrobacterium strain. The use of Agrobacterium shooter mutants or strains carrying an isolated cytokinin gene in order to influence endogenous phytohormone ratios did not result in the formation of shoots nor did it increase levels of regeneration in the first method. Further optimization is in order and in progress.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 3 (1985), S. 61-73 
    ISSN: 1573-5044
    Keywords: plant cell suspension cultures ; callus cultures ; L-DOPA ; catecholamines ; Mucuna pruriens ; Fabaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methods for induction and maintenance of callus as well as cell suspension cultures of Mucuna pruriens are described. The presence of 3- (3, 4-dihy droxyphenyl 1)-L-alanine (L-DOPA) in these cell suspension cultures was demonstrated by means of TLC and HPLC, the latter being equipped with an ultra-violet or a more specific electrochemical detector. Further methods employed were GC, also in combination with mass spectrometry, and thermal desorption chemical ionization mass spectrometry.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 3 (1985), S. 75-82 
    ISSN: 1573-5044
    Keywords: plant cell suspension cultures ; L-DOPA ; catecholamines ; Mucuna pruriens ; Fabaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The endogenous synthesis of 3-(3,4-dihydroxyphenyl)-L-alanine (L-DOPA) by cell suspension cultures of Mucuna pruriens was found to be influenced by several environmental parameters. The nature of the nitrogen source as well as the concentration of nitrogen containing salts, sucrose and phosphate in the culture medium were found to affect the biosynthesis of L-DOPA. Addition of 2, 4-dichlorophenoxyacetic acid to the medium suppressed L-DOPA production; continuous illumination of the cultures had a strong beneficial effect on L-DOPA production. L-DOPA was accumulated intracellularly by the cell suspension cultures. These observations further demonstrate that for certain products of plant cell suspensions product synthesis can be manipulated by a proper selection of specified nutrients.
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