鼠疫菌OxyR调控子蛋白对rovA基因的转录调控研究  

Study on Transcriptional Regulation of rovA by OxyR in Yersinia pestis

作　　者：倪斌[1] 刘磊[2] 黄新祥[1] 韩延平[2] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212013 [2]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《动物医学进展》2013年第12期60-64,共5页Progress In Veterinary Medicine

基　　金：病原微生物生物安全国家重点实验室开放研究基金资助项目(SKLPBS1112);江苏省高校自然科学研究项目(12KJD310001);江苏大学高级专业人才科研启动基金项目(10JDG044)

摘　　要：应用分子生物学技术,开展鼠疫菌调控子蛋白OxyR对rovAR的转录调控机制研究。提取鼠疫菌野生株(WT)和oxyR突变株(ΔoxyR)的总RNA,采用引物延伸试验研究rovAR的转录起始位点,并根据产物的丰度判断OxyR对rovAR的调控关系。构建克隆有rovAR上游启动子区的lacZ重组质粒,将重组质粒转入WT和ΔoxyR中,通过测定β-半乳糖苷酶活性来比较2株重组菌中的rovAR启动子活性,进一步验证OxyR对rovAR的调控关系。进而利用大肠埃希菌OxyR识别基序,预测OxyR对rovAR启动子区的结合位点。发现rovAR的2个转录起始位点均受OxyR的激活,且rovAR启动子区具有可能的OxyR结合位点。推测OxyR能直接结合到rovAR启动子区而激活其转录表达。To stuy the transcriptional regulation mechanism of rovA by OxyR in Yersinia pestis,total bacte-rial RNAs were extracted from the wide-type（WT）strain and the oxyR null mutant（ΔoxyR）.Primer ex-tension assay was employed to detect the promoter activity（the amount of primer extension product）of ro-vA in WT and that in ΔoxyR.The rovA promoter-proximal region was cloned into the pRW50 containing apromoterless lacZ gene.The recombinant LacZ reporter plasmid was transformed into WT and ΔoxyR,respectively,to measure the promoter activity（theβ-Galactosidase activity）of rovA in WT andΔoxyR by using theβ-Galactosidase Enzyme Assay System.And then the E.coli OxyR consensus was carried out to predict the OxyR binding site to rovA promoter region.All the two transcription starts of rovA were acti-vated by OxyR through directly bind to the promoter sequences.OxyR may directly activate the rovA tran-scription in Yersinia pestis.

关 键 词：鼠疫耶尔森菌 OXYR rovAR 转录调控 

分 类 号：Q753[生物学—分子生物学]