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  • 1
    Electronic Resource
    Electronic Resource
    Transformation in the methylotrophic yeast Pichia methanolica utilizing homologous ADE1 and heterologous Saccharomyces cerevisiae ADE2 and LEU2 genes as genetic markers (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 9 (1993), S. 1189-1197 
    Details
    ISSN: 0749-503X
    Keywords: Methylotrophic yeast ; genetic transformation ; non-homologous integration ; gene disruption ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Development of transformation systems for methylotrophic yeasts is the starting point for research aimed at developing molecular genetics of these genera and will be the key to their further successful use in biotechnology. We transformed Pichia methanolica using selector genes ADE2 and LEU2 from Saccharomyces cerevisiae and ADE1 (homologue of S. cerevisiae ADE2 gene) from P. methanolica which was cloned and sequenced in our laboratory (Hiep et al., 1991). Lithium transformation of P. methanolica strains was inefficient with intact plasmids. Linearization of plasmids at a unique restriction site within the ADE1 gene prior to transformation substantially increased its frequency. Transformation with linear ADE1, ADE2 or LEU2 gene fragments was even more effective. Introduced DNA fragments either circularized in vivo, irrespective of the structures of their ends, giving unstable transformants; or integrated at different sites of the host genome. Using this transformation system, we obtained a disruption of the ADE1 gene on the chromosome by inserting the S. cerevisiae LEU2 gene. The disruption mutation ade1::LEU2 was used to study the mechanism of intragenic recombination in P. methanolica.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320091105
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    The 5-aminoimidazole ribonucleotide-carboxylase structural gene of the methylotrophic yeast Pichia methanolica: Cloning, sequencing and homology analysis (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 9 (1993), S. 1251-1258 
    Details
    ISSN: 0749-503X
    Keywords: Methylotrophic yeasts ; phosphoribosyl-5-aminoimidazole-carboxylase ; gene homology analysis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ADE1 gene of the yeast Pichia methanolica encodes phosphoribosyl-5-aminoimidazole-carboxylase (AIRC, EC 4.1.1.21), which is involved in purine biosynthesis. The gene was cloned by complementation of an ade2 mutation in Saccharomyces cerevisiae and a 3077 nucleotide DNA fragment was sequenced. The sequence possessed a single open reading frame, corresponding to a 543 amino acid sequence. The sequence of this putative protein has been compared to the proteins of homologous genes from S. cerevisiae, Schizosaccharomyces pombe, Escherichia coli, chicken and man. The analysis revealed remarkable homology between yeast AIRCs, while for other proteins homology was limited to defined regions.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320091112
    Library Location Call Number Volume/Issue/Year Availability
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    Paper (German National Licenses)
    Fulltext
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