H-NS对副溶血弧菌泳动能力的调控研究  

Regulation of swimming motility by H-NS in Vibrio parahaemolyticus

作　　者：王洁[1] 林磊[1] 孙凤军[3] 董新波[1] 侯书宁 周冬生[1] 殷喆[1] 张义全[2] 

机构地区：[1]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 [2]江苏大学医学院,江苏镇江212013 [3]第三军医大学附属西南医院药剂科,重庆400038

出　　处：《军事医学》2015年第9期694-697,共4页Military Medical Sciences

基　　金：江苏大学高级人才科研启动基金项目(14JDG166);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(SKLPBS1438,SKLPBS1437);国家自然科学基金面上项目(31170127)

摘　　要：目的研究H-NS对副溶血弧菌(Vibrio parahaemolyticus,VP)泳动能力的调控作用。方法将VP接种于泳动实验平板上,37℃静置培养4.5 h后,通过比较不同菌株间菌苔直径的差异来判定H-NS对泳动表型的影响。提取WT和hns基因突变株(Δhns)的总RNA,采用实时定量RT-PCR方法研究H-NS对极鞭毛蛋白基因fla A的转录调控关系。将fla A启动子区DNA克隆入p HRP309质粒的β-半乳糖苷酶基因上游,构建Lac Z重组质粒,并将该重组质粒转入WT和Δhns中,通过比较二者中β-半乳糖苷酶活性的差异研究H-NS对fla A的调控关系。结果与结论动力表型结果显示,H-NS可促进VP的泳动能力;实时定量RT-PCR和Lac Z报告基因融合实验结果表明H-NS正调控fla A的转录。这表明H-NS至少可通过激活fla A的转录而促进VP的泳动能力。Objective To investigate the regulation of swimming motility by H-NS in Vibrio parahaemolyticus（VP）. Methods VP was inoculated into the semi-solid swimming agar plate containing 1% Oxoid tryptone, 2% NaCl, 0.5%Difco Noble Agar, and 0.1% arabinose followed by incubation at 37℃ for 4.5 h before the diameters of bacterial lawns were measured.Total RNAs were extracted from the wild-type （WT） strains and the hns null mutant （Δhns）, and the quantitative real-time（ RT）-PCR（ qRT-PCR） was carried out to calculate the transcriptional variation of flaA between WT andΔhns strains.The entire promoter DNA region of flaA was amplified and cloned into the lacZ fusion vector pHRP309 containing a promoterless lacZ gene. The recombinant lacZ reporter plasmid was transformed into WT and Δhns, respectively, to measure the β-galactosidase activities in cellular extracts using the β-galactosidase enzyme assay system. Results and Conclusion The phenotype results showed that swimming motility of VP was enhanced by H-NS.The qRT-PCR and LacZ fusion results indicated that the transcription of flaA was positively regulated by H-NS.Collectively, H-NS promotes the swimming motility of VP, at least partly, by activating the transcription of flaA.

关 键 词：副溶血弧菌 泳动 

分 类 号：R378.3[医药卫生—病原生物学]