Targeted integration into the Acremonium chrysogenum genome: disruption of the pcbC gene.
作者： Markus Walz , Ulrich Kück , None
DOI: 10.1007/BF00351851
关键词:
摘要: The cephalosporin C-producing fungus Acremonium chrysogenum was transformed to hygromycin B resistance using different vector constructs. These constructs contain sequences of the pcbC gene from A. chrysogenum, encoding isopenicillin N synthetase. Detailed analysis transformants, including pulsed-field gel electrophoresis (PFGE), suggests that integration multiple copies takes place predominantly via non-homologous integration. By increasing length vector-DNA homologous genomic DNA, occurs more frequently into chromosome VI, carrying endogencous copy. In disruption experiments, homology required obtain C-minus transformants investigated. Inactivation observed only when fragments than 3.0 kb were used on both sites cassette. Southern indicated homologous, as well heterologous, recombinant DNA. leads appearance truncated transcripts.
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