用质粒载体转化林肯链霉菌原生质体。

Transformation of Streptomyces lincolnensis protoplasts with plasmid vectors.

作者信息

Jandová Z, Tichý P

机构信息

Institute of Microbiology, Czechoslovak Academy of Sciences, Prague.

出版信息

Folia Microbiol (Praha). 1992;37(3):181-7. doi: 10.1007/BF02933144.

DOI:10.1007/BF02933144

PMID:1505879

Abstract

A method for the preparation and regeneration of protoplasts of Streptomyces lincolnensis is described. Mycelium in the early exponential phase appeared to be most suitable for this purpose and yielded up to 25% regenerated intact cells. Transformation of S. lincolnensis protoplasts was achieved using broad-host-range streptomycete plasmid vectors pIJ622, pMP66, pRS410 and pIJ943 constructed from replicons pIJ101, pSLG33 and SCP2. The efficiency of transformation was 3.10(3) transformants per micrograms plasmid DNA when (2-5).10(7) recipient protoplasts were used. Interspecific transformations showed that there is no efficient restriction system in S. lincolnensis that would limit the transfer of genetic information from S. lividans or E. coli.

摘要

描述了一种制备和再生林肯链霉菌原生质体的方法。指数生长期早期的菌丝体似乎最适合此目的，可产生高达25%的完整再生细胞。使用由复制子pIJ101、pSLG33和SCP2构建的广宿主链霉菌质粒载体pIJ622、pMP66、pRS410和pIJ943实现了林肯链霉菌原生质体的转化。当使用(2 - 5)×10⁷个受体原生质体时，转化效率为每微克质粒DNA有3×10³个转化子。种间转化表明，林肯链霉菌中不存在会限制来自淡紫链霉菌或大肠杆菌的遗传信息转移的有效限制系统。