Inhibition of RecBCD in Klebsiella pneumoniae by Gam and its effect on the efficiency of gene replacement
2016-02
发表期刊JOURNAL OF BASIC MICROBIOLOGY (IF:3.5[JCR-2023],3.3[5-Year])
ISSN0233-111X
卷号56期号:2页码:120-126
发表状态已发表
DOI10.1002/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
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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
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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.
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