阿维链霉菌
生物
互补
质粒
链霉菌
遗传学
基因
同色链霉菌
限制地图
突变体
DNA
分子生物学
细菌
作者
Douglas J. MacNeil,Keith M. Gewain,Carolyn L. Ruby,Gabe Dezeny,Patrice H. Gibbons,Tanya MacNeil
出处
期刊:Gene
[Elsevier]
日期:1992-02-01
卷期号:111 (1): 61-68
被引量:668
标识
DOI:10.1016/0378-1119(92)90603-m
摘要
An integration vector for gene analysis in Streptomyces has been constructed. This vector replicates in Escherichia coli, and integrates into Streptomyces by homologous recombination between a cloned fragment and the genome. To overcome methylation-specific restriction barriers, an E. coli mutant triply defective in DNA methylation was constructed as a source for the integration plasmids. The frequency of integration of pVE616 derivatives into the Streptomyces avermitilis genome was proportional to the size of the cloned DNA. Derivatives of pVE616, containing fragments from pVE650, a plsmid with a 24-kb insert of S. avermitilis DNA, were used in complementation analyses of seven S. avermitilis mutants defective in glycosylation of avermectin (Av). Three complementation groups, located in a 7-kb region, were identified. Derivatives of pVE616, containing fragments from the 18-kb of DNA adjacent to the glycosylation region, were integrated into an Av producer. Av produced from the integrants was substantially reduced, indicating that the 18 kb encodes gene products which are involved in Av biosynthesis.
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