军事医学
軍事醫學
군사의학
Military Medical Sciences
2015年
9期
694-697
,共4页
王洁%林磊%孙凤军%董新波%侯书宁%周冬生%殷喆%张义全
王潔%林磊%孫鳳軍%董新波%侯書寧%週鼕生%慇喆%張義全
왕길%림뢰%손봉군%동신파%후서저%주동생%은철%장의전
副溶血弧菌%H-NS%泳动%flaA
副溶血弧菌%H-NS%泳動%flaA
부용혈호균%H-NS%영동%flaA
Vibrio parahaemolyticus%H-NS%swimming motility%flaA
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flaA的转录调控关系。将flaA启动子区DNA克隆入pHRP309质粒的β-半乳糖苷酶基因上游，构建LacZ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flaA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LacZ报告基因融合实验结果表明H-NS正调控flaA的转录。这表明H-NS至少可通过激活flaA的转录而促进VP的泳动能力。
目的：研究H-NS對副溶血弧菌（ Vibrio parahaemolyticus，VP）泳動能力的調控作用。方法將VP接種于泳動實驗平闆上，37℃靜置培養4．5 h後，通過比較不同菌株間菌苔直徑的差異來判定H-NS對泳動錶型的影響。提取WT和hns基因突變株（Δhns）的總RNA，採用實時定量RT-PCR方法研究H-NS對極鞭毛蛋白基因flaA的轉錄調控關繫。將flaA啟動子區DNA剋隆入pHRP309質粒的β-半乳糖苷酶基因上遊，構建LacZ重組質粒，併將該重組質粒轉入WT和Δhns中，通過比較二者中β-半乳糖苷酶活性的差異研究H-NS對flaA的調控關繫。結果與結論動力錶型結果顯示，H-NS可促進VP的泳動能力；實時定量RT-PCR和LacZ報告基因融閤實驗結果錶明H-NS正調控flaA的轉錄。這錶明H-NS至少可通過激活flaA的轉錄而促進VP的泳動能力。
목적：연구H-NS대부용혈호균（ Vibrio parahaemolyticus，VP）영동능력적조공작용。방법장VP접충우영동실험평판상，37℃정치배양4．5 h후，통과비교불동균주간균태직경적차이래판정H-NS대영동표형적영향。제취WT화hns기인돌변주（Δhns）적총RNA，채용실시정량RT-PCR방법연구H-NS대겁편모단백기인flaA적전록조공관계。장flaA계동자구DNA극륭입pHRP309질립적β-반유당감매기인상유，구건LacZ중조질립，병장해중조질립전입WT화Δhns중，통과비교이자중β-반유당감매활성적차이연구H-NS대flaA적조공관계。결과여결론동력표형결과현시，H-NS가촉진VP적영동능력；실시정량RT-PCR화LacZ보고기인융합실험결과표명H-NS정조공flaA적전록。저표명H-NS지소가통과격활flaA적전록이촉진VP적영동능력。
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.
