[大肠杆菌与链霉菌之间高效接合质粒的构建]

[Construction of efficient conjugal plasmids between Escherichia coli and Streptomycetes].

作者信息

Mo Hong-Bo, Bai Lin-Quan, Wang Sheng-Lan, Yang Ke-Qian

机构信息

Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100080, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2004 Sep;20(5):662-6.

PMID:15973986

Abstract

Conjugal plasmid pGH112 has been developed based on the replicons of Streptomyces coelicolor plasmid SCP2 and E. coli ColE. The plasmid contains ampicilin resistance gene(amp) for selection in E. coli and thiostrepton resistance gene (tsr) for selection in Streptomycetes, and a 0.76 kb oriT fragment of (IncP) RK2. Conjugal transfer of pGH112 was performed from E. coli to S. coelicolor A3(2), S. avermitilis, S. lividans TK54, S. toxytricini NNRL15443, S. venezuelae ISP5230 and Sacc. erythraea by conjugation, results show that the plasmid was able to transfer efficenctly from E. coli to Streptomycetes, was stably inherited in the recipients. pGH113 was constructed from pGH112 by combining the constitutive ermE promoter with green fluorescent protein gene(gfp).

摘要

接合质粒pGH112是基于天蓝色链霉菌质粒SCP2和大肠杆菌ColE的复制子构建的。该质粒含有用于在大肠杆菌中筛选的氨苄青霉素抗性基因（amp）和用于在链霉菌中筛选的硫链丝菌素抗性基因（tsr），以及（IncP）RK2的一个0.76 kb的oriT片段。通过接合作用将pGH112从大肠杆菌转移至天蓝色链霉菌A3(2)、阿维链霉菌、变铅青链霉菌TK54、毒三素链霉菌NNRL15443、委内瑞拉链霉菌ISP5230和红霉菌属，结果表明该质粒能够有效地从大肠杆菌转移至链霉菌，并在受体中稳定遗传。pGH113是通过将组成型ermE启动子与绿色荧光蛋白基因（gfp）相结合，从pGH112构建而来的。