用于链霉菌属的多功能高效接合质粒的开发

Development of a multifunctional and efficient conjugal plasmid for use in Streptomyces spp.

作者信息

Mo H B, Chen X Y, Liu Y, Xia F, Yang K Q

机构信息

Center of Biochemistry and Molecular Biology [corrected] College of Medical Sciences, Jinan University [corrected] Guangzhou, 510632, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2006 May;70(6):705-10. doi: 10.1007/s00253-006-0324-7. Epub 2006 Mar 11.

DOI:10.1007/s00253-006-0324-7

PMID:16532313

Abstract

A plasmid, pGB112, has recently been developed to transfer DNA from Escherichia coli to Streptomyces spp via conjugation. This technique made use of (A) E. coli replicon, (B) ampicillin (amp) resistance gene for selection in E. coli and thiostrepton (tsr) resistance gene for selection in Streptomyces, (C) a fragment of SCP2* replicon, (D) a 2.6 kb fragment of tra-cassette which consists of pIJ101 transfer gene (tra) and two ermE promoters, (E) a 0.8 kb fragment of oriT of (IncP) RK2. The results showed that this plasmid was able to transfer plasmid DNA from E. coli to Streptomyces coelicolor via conjugation, and that it could also transfer DNA between Streptomyces strains. Since this plasmid has both pBR322 and SCP2* replicons, it may provide a novel and useful method for genetic operation in E. coli and Streptomyces.

摘要

最近开发了一种质粒pGB112，用于通过接合作用将DNA从大肠杆菌转移到链霉菌属。该技术利用了：（A）大肠杆菌复制子；（B）用于在大肠杆菌中筛选的氨苄青霉素（amp）抗性基因和用于在链霉菌中筛选的硫链丝菌素（tsr）抗性基因；（C）SCP2复制子的一个片段；（D）一个2.6 kb的tra盒片段，其由pIJ101转移基因（tra）和两个ermE启动子组成；（E）（IncP）RK2的oriT的一个0.8 kb片段。结果表明，该质粒能够通过接合作用将质粒DNA从大肠杆菌转移到天蓝色链霉菌，并且还能在链霉菌菌株之间转移DNA。由于该质粒同时具有pBR322和SCP2复制子，它可能为大肠杆菌和链霉菌的基因操作提供一种新颖且有用的方法。