Targeted Chromosomal Knockouts in Mycoplasma pneumoniae

被引：25

作者：

Krishnakumar, Radha ^{[1
]}

Assad-Garcia, Nacyra ^{[1
]}

Benders, Gwynedd A. ^{[2
]}

Phan, Quang ^{[1
]}

Montague, Michael G. ^{[1
]}

Glass, John I. ^{[1
]}

机构：

[1] J Craig Venter Inst, Rockville, MD 20850 USA

[2] J Craig Venter Inst, San Diego, CA 92121 USA

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2010年 / 76卷 / 15期

关键词：

HOMOLOGOUS RECOMBINATION; VIRULENCE DETERMINANT; CONSTRUCTION; GENOME; GENITALIUM; BACTERIUM; GALLISEPTICUM; DERIVATIVES; DISRUPTION; AGALACTIAE;

D O I：

10.1128/AEM.00024-10

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：

Most gene knockouts in mycoplasmas are achieved through labor-intensive transposon mutagenesis. Here, we describe a method for making targeted deletions in Mycoplasma pneumoniae by use of homologous recombination. In this method, M. pneumoniae is transformed with a plasmid carrying an antibiotic resistance marker flanked by 1-kb regions surrounding the target gene. Following selection for the antibiotic resistance, colonies are screened for double crossovers which indicate complete deletion of the target open reading frame.

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收藏

页码：5297 / 5299

页数：3

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