High-throughput cloning and expression in recalcitrant bacteria

被引：85

作者：

Geertsma, Eric R.

Poolman, Bert

机构：

[1] Univ Groningen, Dept Biochem, Groningen Biomol Sci & Biotechnol Inst, NL-9747 AG Groningen, Netherlands

[2] Univ Groningen, Zernike Inst Adv Mat, Dept Biochem, NL-9747 AG Groningen, Netherlands

来源：

NATURE METHODS | 2007年 / 4卷 / 09期

关键词：

D O I：

10.1038/NMETH1073

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We developed a generic method for high-throughput cloning in bacteria that are less amenable to conventional DNA manipulations. The method involves ligation-independent cloning in an intermediary Escherichia coli vector, which is rapidly converted via vector-backbone exchange (VBEx) into an organism-specific plasmid ready for high-efficiency transformation. We demonstrated VBEx proof of principle for Lactococcus lactis, but the method can be adapted to all organisms for which plasmids are available.

引用

页码：705 / 707

页数：3

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