Genetic tools for the piezophilic hyperthermophilic archaeon Pyrococcus yayanosii
Li, Xuegong1,2; Fu, Ling1; Li, Zhen1; Ma, Xiaopan1; Xiao, Xiang1,3,4; Xu, Jun1,3,4
2015
发表期刊EXTREMOPHILES
卷号19期号:1页码:59-67
摘要The hyperthermophile Pyrococcus yayanosii CH1 is the only high-pressure-requiring microorganism obtained thus far within the archaea domain or among all non-psychrophiles in any domain. In this study, we developed a genetic manipulation system for P. yayanosii after first isolating a facultatively piezophilic derivative strain, designated P. yayanosii A1. The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase gene was overexpressed in strain P. yayanosii A1 and was demonstrated to confer host cell resistance against simvastatin. Furthermore, using simvastatin as a selection marker, the endogenous pyrF of P. yayanosii A1 was disrupted through homologous recombination, thus generating the additional host strain P. yayanosii A2 (Delta pyrF). A markerless gene disruption vector was constructed by incorporating a pyrF-sim (R) cassette that enables the combined use of simvastatin resistance for positive selection and 5-FOA for counter selection. The utility of this versatile disruption system was demonstrated by deleting the carbon-nitrogen hydrolase of P. yayanosii strain A1. These results demonstrate that a variety of genetic tools are now in place to study unknown gene function and the molecular mechanisms of piezophilic adaptation in P. yayanosii.
文章类型Article
关键词Piezophile Genetic Tool Pyrococcus Yayanosii Hyperthermophilic Archaeon
学科领域海洋生物学
WOS标题词Science & Technology ; Life Sciences & Biomedicine
DOI10.1007/s00792-014-0705-2
URL查看原文
收录类别SCI
语种英语
关键词[WOS]Piezophile ; Genetic tool ; Pyrococcus yayanosii ; Hyperthermophilic archaeon
WOS类目Biochemistry & Molecular Biology ; Microbiology
WOS记录号WOS:000346657500006
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被引频次:6[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.idsse.ac.cn/handle/183446/1204
专题深海科学研究部_深海生物学研究室_深海微生物细胞生物学研究组
作者单位1.Shanghai Jiao Tong Univ, Inst Oceanol, Shanghai 200240, Peoples R China
2.Chinese Acad Sci, Sanya Inst Deep Sea Sci & Engn, Dept Deep Sea Sci, Sanya 572000, Peoples R China
3.Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China
4.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China
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Li, Xuegong,Fu, Ling,Li, Zhen,et al. Genetic tools for the piezophilic hyperthermophilic archaeon Pyrococcus yayanosii[J]. EXTREMOPHILES,2015,19(1):59-67.
APA Li, Xuegong,Fu, Ling,Li, Zhen,Ma, Xiaopan,Xiao, Xiang,&Xu, Jun.(2015).Genetic tools for the piezophilic hyperthermophilic archaeon Pyrococcus yayanosii.EXTREMOPHILES,19(1),59-67.
MLA Li, Xuegong,et al."Genetic tools for the piezophilic hyperthermophilic archaeon Pyrococcus yayanosii".EXTREMOPHILES 19.1(2015):59-67.
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