ISSN:
1432-0983
Keywords:
Acremonium chrysogenum
;
pcbC gene
;
Transformation
;
Gene disruption
;
Pulsed-field gel electrophoresis
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract 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 of transformants, including pulsed-field gel electrophoresis (PFGE), suggests that integration of multiple vector copies takes place predominantly via non-homologous integration. By increasing the length of vector-DNA homologous to genomic DNA, integration occurs more frequently into chromosome VI, carrying the endogencous pcbC gene copy. In gene disruption experiments, the length of vector homology required to obtain cephalosporin C-minus transformants was investigated. Inactivation of the pcbC gene was observed only when homologous fragments of more than 3.0 kb were used on both sites of the resistance cassette. Southern analysis indicated homologous, as well as heterologous, integration of recombinant DNA. The integration of multiple vector copies leads to the appearance of truncated pcbC transcripts.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00351851
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