Inhibition of RecBCD in Klebsiella pneumoniae by Gam and its effect on the efficiency of gene replacement

doi:10.1002/jobm.201400953

	Inhibition of RecBCD in Klebsiella pneumoniae by Gam and its effect on the efficiency of gene replacement
	Chen, Chuan 1,2; Wei, Dong 1; Liu, Pengfu 1; Wang, Min 2; Shi, Jiping1,3 ; Jiang, Biao 1; Hao, Jian 1
	2016-02
发表期刊	JOURNAL OF BASIC MICROBIOLOGY (IF:3.5[JCR-2023],3.3[5-Year])
ISSN	0233-111X
卷号	56 期号:2 页码:120-126
发表状态	已发表
DOI	10.1002/jobm.201400953
摘要	Gam protein is an inhibitor of the host RecBCD exonuclease, and this inhibition is essential to the proficiency of Red recombinase-mediated gene replacement. In Klebsiella pneumoniae, the efficiency of this gene replacement was lower than that in Escherichia coli, and the minimum length of homologous extensions required was longer. Thus, it was supposed that the inhibitory effect of Gam against RecBCD was weak in K. pneumoniae. To test this hypothesis, a Gam-deficient Red recombinase expression plasmid and a recB K. pneumoniae mutant were constructed. The Gam-deficient Red recombinase showed a reduced capacity for gene replacement compared with that of the complete Red recombinase. The efficiency of gene replacement in the recB mutant was 6-8 times higher than the wild-type strain, and the minimum length for the homologous extensions was reduced to 100bp. These results indicate that Gam does inhibit the RecBCD exonuclease in K. pneumoniae, but that this inhibition is not stringent. Furthermore, mutation of recB presents a convenient and efficient method to enhance the Red recombinase assisted gene replacement in K. pneumoniae.
关键词	Gene replacement Klebsiella pneumoniae RecBCD Red recombinase Gam
收录类别	SCI
语种	英语
资助项目	National Natural Science Foundation of China[20906076]
WOS研究方向	Microbiology
WOS类目	Microbiology
WOS记录号	WOS:000369847800002
出版者	WILEY
WOS关键词	LAMBDA-RED ; ESCHERICHIA-COLI ; CHROMOSOMAL GENES ; PCR PRODUCTS ; SYSTEM ; 1,3-PROPANEDIOL ; INACTIVATION ; EXPRESSION ; MECHANISM ; PLASMIDS
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1930
专题	生命科学与技术学院_特聘教授组_史吉平组
通讯作者	Shi, Jiping; Hao, Jian
作者单位	1.Chinese Acad Sci, Shanghai Adv Res Inst, Lab Biorefinery, 99 Haike Rd, Shanghai 201210, Peoples R China 2.Tianjin Univ Sci & Technol, Coll Biotechnol, Key Lab Ind Fermentat Microbiol, Minist Educ, Tianjin, Peoples R China 3.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Chen, Chuan,Wei, Dong,Liu, Pengfu,et al. Inhibition of RecBCD in Klebsiella pneumoniae by Gam and its effect on the efficiency of gene replacement[J]. JOURNAL OF BASIC MICROBIOLOGY,2016,56(2):120-126.
APA	Chen, Chuan.,Wei, Dong.,Liu, Pengfu.,Wang, Min.,Shi, Jiping.,...&Hao, Jian.(2016).Inhibition of RecBCD in Klebsiella pneumoniae by Gam and its effect on the efficiency of gene replacement.JOURNAL OF BASIC MICROBIOLOGY,56(2),120-126.
MLA	Chen, Chuan,et al."Inhibition of RecBCD in Klebsiella pneumoniae by Gam and its effect on the efficiency of gene replacement".JOURNAL OF BASIC MICROBIOLOGY 56.2(2016):120-126.