Targeted mutagenesis of Burkholderia thailandensis and Burkholdetia pseudomallei through natural transformation of PCR fragments

被引：49

作者：

Thongdee, Metawee ^{[1
,2
]}

Gallagher, Larry A. ^{[1
]}

Schell, Mark ^{[3
]}

Dharakul, Tararaj ^{[2
]}

Songsivilai, Sirirurg ^{[2
]}

Manoil, Colin ^{[1
]}

机构：

[1] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA

[2] Mahidol Univ, Siriraj Hosp, Fac Med, Dept Immunol, Bangkok 10700, Thailand

[3] Univ Georgia, Dept Microbiol, Athens, GA 30602 USA

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2008年 / 74卷 / 10期

关键词：

D O I：

10.1128/AEM.00030-08

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Burkholderia pseudomallei is the causative agent of melioidosis, an overwhelming, rapidly fatal septic infection, and B. thailandensis is a closely related, less virulent species. Both organisms are naturally competent for DNA transformation, and this report describes a procedure exploiting this property for the rapid generation of marked deletion mutations by using PCR products. The method was employed to create 61 mutant strains. Several selectable elements were employed, including elements carrying loxP and FRT recombinase recognition sites to facilitate resistance marker excision. Chromosomal mutations could also be transferred readily between strains by transformation. The availability of simple procedures for creating defined chromosomal mutations and moving them between strains should facilitate genetic analysis of virulence and other traits of these two Burkholderia species.

引用

页码：2985 / 2989

页数：5

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