用于链霉菌属和大肠杆菌的两种稳定双功能质粒的构建

Construction of two stable bifunctional plasmids for Streptomyces spp. and Escherichia coli.

作者信息

Durajlija S, Pigac J, Gamulin V

机构信息

Department of Organic Chemistry and Biochemistry, Ruder Bosković Institute, University of Zagreb, Yugoslavia.

出版信息

FEMS Microbiol Lett. 1991 Oct 15;67(3):317-21. doi: 10.1016/0378-1097(91)90495-v.

DOI:10.1016/0378-1097(91)90495-v

PMID:1769540

Abstract

Two bifunctional plasmid vectors pZG5 (7.45 kb) and pZG6 (6.95 kb), for gene transfer between Streptomyces spp. and Escherichia coli have been constructed by fusion of the multicopy broad-host-range Streptomyces plasmid pIJ350 with E. coli plasmids Bluescribe M13- (pZG5) or pUC18 (pZG6). Both plasmids possess several unique restriction sites suitable for DNA cloning. Stable transformants of Streptomyces rimosus R6 and S. lividans 66 were obtained, harboring intact plasmids regardless of colony age or multiple subculturing. Moreover, pZG5 and pZG6 were successfully used to introduce several homologous transfer RNA genes into S. rimosus.

摘要

通过将多拷贝广宿主链霉菌质粒pIJ350与大肠杆菌质粒Bluescribe M13-（pZG5）或pUC18（pZG6）融合，构建了两种用于链霉菌属和大肠杆菌之间基因转移的双功能质粒载体pZG5（7.45 kb）和pZG6（6.95 kb）。这两种质粒都具有几个适合DNA克隆的独特限制酶切位点。获得了龟裂链霉菌R6和变铅青链霉菌66的稳定转化体，无论菌落年龄或多次传代，它们都含有完整的质粒。此外，pZG5和pZG6已成功用于将几个同源转移RNA基因导入龟裂链霉菌。