期刊
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
卷 73, 期 8, 页码 2735-2743出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.02829-06
关键词
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资金
- NIGMS NIH HHS [R01 GM037949, GM37949] Funding Source: Medline
Mobile group II introns (targetrons) can be programmed for insertion into virtually any desired DNA target with high frequency and specificity. Here, we show that targetrons expressed via an m-toluic acid-inducible promoter from a broad-host-range vector containing an RK2 minireplicon can be used for efficient gene targeting in a variety of gram-negative bacteria, including Escherichia coli, Pseudomonas aeruginosa, and Agrobacterium tumefaciens. Targetrons expressed from donor plasmids introduced by electroporation or conjugation yielded targeted disruptions at frequencies of 1 to 58% of screened colonies in the E. coli lacZ, P. aeruginosa pqsA and pqsH, and A. tumefaciens aopB and chvI genes. The development of this broad-host-range system for targetron expression should facilitate gene targeting in many bacteria.
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