Teth137, a Conserved Factor of Unknown Function from Thermoanaerobacter ethanolicus JW200, Represses the Transcription of the adhE Gene In Vitro

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	Teth137, a Conserved Factor of Unknown Function from Thermoanaerobacter ethanolicus JW200, Represses the Transcription of the adhE Gene In Vitro
	Jing, Qingqing 1; Wang, Jingkai 2; Wu, Guogan 2,3
刊名	INDIAN JOURNAL OF MICROBIOLOGY
	2013-06-01
卷号	53 期号:2 页码:149-154
关键词	Thermoanaerobacter ethanolicus AdhE GEMSA In vitro transcription
英文摘要	Teth137, a 13.7 kD protein of unknown function from Thermoanaerobacter ethanolicus JW200, is encoded by 360 nucleotides and has been obtained by DNA-coupled column previously. However, no function study of Teth137 has been published. Homologous modeling of Teth137 shows the protein is comprised of a helix-turn-helix motif which is a typical DNA-binding domain. Therefore, it is speculated Teth137 is a DNA-binding protein and involved in transcription of the adhE gene (encodes alcohol dehydrogenase E). To investigate the function of Teth137, recombinant Teth137 is overexpressed in Escherichia coli JM109 and purified by DEAE column. Purified Teth137 exhibits the affinity with the adhE promoter region in gel electrophoresis mobility shift assay (GEMSA). Teth137 at the concentration of 48 mu M retards the migration of 5 nM of probe in the presence of the competitor DNA. Mutant analysis indicates that S69, T70, P71 and T72 are critical to protein-DNA interface; Gly substitutions at these residues results in the loss of the binding ability with the adhE promoter region. Moreover, T. ethanolicus JW200 RNA polymerase, sigma subunit and template plasmid are prepared for in vitro transcription assay to detect the regulation function of Teth137. The results of the in vitro transcription show that the transcription of 5 nM of the template plasmid is inhibited by 48 mu M of Teth137.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology ; Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology ; Microbiology
关键词[WOS]	SECONDARY-ALCOHOL DEHYDROGENASE ; RNA-POLYMERASE ; PURIFICATION ; EXPRESSION ; CLONING ; 39E
收录类别	SCI
语种	英语
WOS记录号	WOS:000317693300005
公开日期	2015-12-24
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn/handle/337004/6046]
专题	青岛生物能源与过程研究所_仿真模拟团队
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Modeling & Simulat Grp, Qingdao 266061, Peoples R China 2.Nanjing Normal Univ, Coll Life Sci, Jiangsu Key Lab Biodivers & Bioresources, Nanjing 210046, Jiangsu, Peoples R China 3.Southeast Univ, Coll Chem & Chem Engn, Nanjing 210096, Jiangsu, Peoples R China
推荐引用方式 GB/T 7714	Jing, Qingqing,Wang, Jingkai,Wu, Guogan. Teth137, a Conserved Factor of Unknown Function from Thermoanaerobacter ethanolicus JW200, Represses the Transcription of the adhE Gene In Vitro[J]. INDIAN JOURNAL OF MICROBIOLOGY,2013,53(2):149-154.
APA	Jing, Qingqing,Wang, Jingkai,&Wu, Guogan.(2013).Teth137, a Conserved Factor of Unknown Function from Thermoanaerobacter ethanolicus JW200, Represses the Transcription of the adhE Gene In Vitro.INDIAN JOURNAL OF MICROBIOLOGY,53(2),149-154.
MLA	Jing, Qingqing,et al."Teth137, a Conserved Factor of Unknown Function from Thermoanaerobacter ethanolicus JW200, Represses the Transcription of the adhE Gene In Vitro".INDIAN JOURNAL OF MICROBIOLOGY 53.2(2013):149-154.