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ShanghaiTech University Knowledge Management System
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]) |
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...
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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 |
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. |
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