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  • Aspergillus niger  (1)
  • genetic analysis  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Expression of the glucose oxidase gene from Aspergillus niger in Hansenula polymorpha and its use as a reporter gene to isolate regulatory mutations (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 9 (1993), S. 625-635 
    Details
    ISSN: 0749-503X
    Keywords: Aspergillus niger ; gene expression ; glucose oxidase ; Hansenula polymorpha ; MF-α secretion leader sequence ; methylotrophic yeast ; methanol oxidase promoter ; recombinant protein ; regulatory mutants ; reporter gene ; super-secretion mutant ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The glucose oxidase gene (god) from Aspergillus niger was expressed in Hansenula polymorpha using the methanol oxidase promoter and transcription termination region and the MF-α leader sequence from Saccharomyces cerevisiae to direct secretion. The expression cassette was cloned into the S. cerevisiae vector YEp13 and used to transform H. polymorpha strain A16. In the initial transformants plasmid replication was unstable, but was stabilized by a growth regime consisting of alternating cycles of selective and non-selective growth. The stabilized strain was grown to high cell density by fed-batch fermentation. Upon induction of the MOX promoter, glucose oxidase synthesis was initiated. At the end of the fermentation, the culture density was 76 g dry weight/1 and 108 IU/ml (0·5 g/l or 0·65% dry weight) glucose oxidase was found in the culture medium; a further 86 IU/ml (0·43 g/l or 0·56% dry weight) was recovered from the cell lysate. A plate assay was used to monitor glucose oxidase levels in individual colonies. This was then used to isolate mutants which showed abnormal regulation of god expression or which showed an altered pattern of secretion. One mutant, which showed increased production of glucose oxidase, was grown to high cell density by fed-batch fermentation (100·6 g/l) and produced 445 IU/ml (2·25 g/l or 2·2% dry weight) extracellularly and 76 IU/ml (0·38 g/l or 0·4% dry weight) intracellularly. The mutant thus not only increased total production but exported 83% of the total enzyme made compared to 55% in the parent strain.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320090609
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  • 2
    Electronic Resource
    Electronic Resource
    Genetic analysis in the methylotrophic yeast Hansenula polymorpha (1988)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 4 (1988), S. 293-303 
    Details
    ISSN: 0749-503X
    Keywords: Hansenula polymorpha ; methylotrophic yeast ; genetic analysis ; methanol mutant ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Techniques are described for the induction, isolation, and characterization of mutants of Hansenula polymorpha. In addition, techniques for controlled passage through the life cycle and genetic analyses, including complementation, tetrad and random spore analysis, have been developed and used to assign mutants to 62 complementation groups. We report that organism conforms to the expected genetics of a homothallic yeast and displays a Mendelian segregation of genes through meiosis. Preliminary mapping data are presented indicating linkage of three genes on a single linkage fragment. Enymatic analysis of methanol-non-utilizing mutants identified one class which is totally deficient in the key assimilatroy enzyme, dihydroxyacetone synthase.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320040407
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    Paper (German National Licenses)
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