A new theta-type thermosensitive replicon from Lactococcus lactis as an integration vector for Enterococcus faecalis

被引：8

作者：

Frère, J ^{[1
]}

Benachour, A

Giard, JC

Laplace, JM

Flahaut, S

Auffray, Y

机构：

[1] Univ Caen, IRBA, Lab Microbiol Environm, F-14032 Caen, France

[2] Univ Fribourg, Inst Biochim, CH-1700 Fribourg, Switzerland

[3] Univ S Alabama, Dept Immunol & Microbiol, Mobile, AL 36688 USA

[4] Univ Littoral Cote dOpale, Biol Cellulaire & Mol Lab, F-62228 Calais, France

来源：

FEMS MICROBIOLOGY LETTERS | 1998年 / 161卷 / 01期

关键词：

thermosensitive replicon; Lactococcus lactis; Enterococcus faecalis; theta replication; integration vector;

D O I：

10.1016/S0378-1097(98)00056-1

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

We isolated a replication thermosensitive mutant of the theta-type lactococcal pUCL22 replicon. An improved version of this thermosensitive replicon was obtained by fusioning the replication repA gene with the downstream repB gene. The resulting plasmid was named pUCB3522Ts. It is highly instable at 42 degrees C in Enterococcus faecalis. Integration into the chromosome via homologous recombination was monitored using the npr gene of E. faecalis JH2-2 as a target. A 513 bp PCR amplification product from an internal region of this npr gene was cloned into pUCB3522Ts. Integration of this construction into the JH2-2 npr gene was selected by shift temperature, from 30 degrees C to 42 degrees C. 85% of the analysed clones showed integration into the npr gene, demonstrating the practicality of this thermosensitive replicon as a genetic integrative tool for E. faecalis. (C) 1998 Published by Elsevier Science B.V.

引用

页码：107 / 114

页数：8

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