Allelic replacement in Staphylococcus aureus with inducible counter-selection

被引：506

作者：

Bae, T ^{[1
]}

Schneewind, O ^{[1
]}

机构：

[1] Univ Chicago, Dept Microbiol, Chicago, IL 60637 USA

来源：

PLASMID | 2006年 / 55卷 / 01期

关键词：

allelic replacement; Staphylococcus aureus; antisense RNA; phage lambda recombination; secY; rocA;

D O I：

10.1016/j.plasmid.2005.05.005

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A method for rapid selection of allelic replacement mutations in the chromosome of Staphylococcus aureus is described. Plasmid pKOR1, an Escherichia colilS. aureus shuttle vector, permits rapid cloning via lambda recombination and ccdB selection. Plasmid transformation of staphylococci and growth at 43 degrees C, a non-permissive condition for pKOR1 replication, selects for homologous recombination and pKOR1 integration into the bacterial chromosome. Anhydrotetracycline-mediated induction of pKOR1-encoded secY antisense transcripts via the Pxyl/tetO promoter, a condition that is not compatible with staphylococcal growth, selects for chromosomal excision and loss of plasmid. Using this strategy, allelic replacements in S. aureus rocA were generated at frequencies that obviated the need for antibiotic marker selection. (c) 2005 Elsevier Inc. All rights reserved.

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页码：58 / 63

页数：6

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