鼠伤寒沙门菌sRNA GcvB和伴侣蛋白Hfq对Ⅲ型分泌系统相关基因的调控  

Regulation of sRNA GcvB and chaperone protein Hfq of Salmonella typhimurium on genes related to typeⅢsecretory system

作　　者：张家莉 潘永 段世宇 令狐远凤 郑如雯 杨琦[1,3] ZHANG Jiali;PAN Yong;DUAN Shiyu;LINGHU Yuanfeng;ZHENG Ruwen;YANG Qi(College of Animal Science,Guizhou University;Research Institute of Animal Husbandry and Veterinary Medicine,Guizhou Academy of Agricultural Sciences;Guizhou Provincial Institute of Animal Diseases,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州大学动物科学学院 [2]贵州省农业科学院畜牧兽医研究所 [3]贵州省动物疫病研究所,贵州贵阳550025

出　　处：《福建农林大学学报（自然科学版）》2023年第4期512-518,共7页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金项目(31760740、31602065)。

摘　　要：细菌的Ⅲ型分泌系统(T3SS)与毒力密切相关。为探究鼠伤寒沙门菌sRNA GcvB和伴侣蛋白Hfq对T3SS相关基因spaP、invC、invG、prgH、prgI、prgJ、prgK的调控,利用Red同源重组技术构建了上述基因与lacZ的融合菌株,在此基础上利用P22噬菌体转导技术分别构建上述融合菌株与gcvB、hfq的单缺失菌株,通过qPCR和β-半乳糖苷酶试验分别检测gcvB、hfq缺失后各基因的转录水平和蛋白水平的变化。结果显示:与标准菌株相比,gcvB缺失后,spaP、invC、prgH、prgI、invG的转录水平和蛋白水平均上调;prgJ、prgK的转录水平上调,蛋白水平下调。hfq缺失后,spaP、invC、prgH、prgI、invG的转录水平和蛋白水平均上调,prgH、prgI、prgJ、prgK的蛋白水平下调。表明Hfq和GcvB均可对spaP、invC、invG、prgH、prgI、prgJ、prgK进行调控。上述结果可为鼠伤寒沙门菌sRNA GcvB和Hfq的作用机理研究提供参考。TypeⅢsecretory system(T3SS)of bacteria is closely related to virulence.To explore the regulation of Salmonella typhi-murium sRNA Gcv B and chaperone protein Hfq on T3SS related genes(spa P,inv C,inv G,prg H,prg I,prg J and prg K),fusion strains of these genes with lac Z gene were constructed by Red homologous recombination technology,followed by the knockout of ei-ther gcv B or hfq gene by P22 phage transduction.q PCR assay andβ-galactosidase assay were performed on the fusion strains to de-tect the effect of the deletion of gcv B or hfq on T3SS related genes at the transcriptional and protein levels.The results showed that the deletion of gcv B up-regulated the transcription and protein levels of spa P,inv C,prg H,prg I and inv G,and up-regulated prg J and prg K at the transcriptional level but down-regulated at the protein level.On the other hand,the transcription and protein levels of spa P,inv C,prg H,prg I and inv G were up-regulated after the deletion of hfq,while the protein levels of prg H,prg I,prg J and prg K were down-regulated.These results indicated that both Hfq and Gcv B can regulate spa P,inv C,inv G,prg H,prg I,prg J and prg K genes.This study laid a theoretical foundation for revealing the functional mechanism of Gcv B and Hfq on S.typhimurium T3SS.

关 键 词：鼠伤寒沙门菌 RED同源重组 GcvB HFQ Ⅲ型分泌系统 

分 类 号：S852.61[农业科学—基础兽医学]