sRNA GcvB对鼠伤寒沙门菌毒力的调控作用研究  

The Regulatory Roles of sRNA GcvB in Virulence in Salmonella typhimurium

作　　者：马忠梅 李能秀 焦健 左雨霏 令一汕 才学鹏[2] 孟庆玲[1] 乔军[1] MA Zhongmei;LI Nengxiu;JIAO Jian;ZUO Yufei;LING Yishan;CAI Xuepeng;MENG Qingling;QIAO Jun(College of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang 832003;China Institute of Veterinary Drug Control,Beijing 730046)

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]中国兽医药品监察所,北京730046

出　　处：《核农学报》2024年第6期1048-1056,共9页Journal of Nuclear Agricultural Sciences

基　　金：国家重点研发计划项目(2016YFD0500900);新疆生产建设兵团国际科技合作计划项目(2019BC004),2023年兵团研究生创新项目。

摘　　要：小RNA(sRNA)是一种调节分子,可以在转录后水平调节基因表达,从而改变细菌的生理特征。为了研究sRNA GcvB在鼠伤寒沙门菌毒力中的调控作用,本研究构建了GcvB基因缺失株、互补株及示踪菌,检测其对鼠伤寒沙门菌生物被膜形成及泳动能力的影响;分析亲本株与缺失株在毒力及感染动力学上的差异,预测并分析了GcvB调控的潜在靶基因,探究了GcvB对ST生物被膜、氧化应激和毒力的调控作用。结果显示,与亲本株和回补株相比,GcvB缺失后生物被膜形成能力升高、抗氧化应激能力显著增强;半数致死量(LD50)上升至3.98倍。实时荧光定量PCR(qRT-PCR)检测靶基因gcvA、bapA、hilA、igaA及sitD,结果显示gcvA、bapA、igaA及sitD的转录水平表达量上调,而hilA下降。本研究证实sRNA GcvB参与了鼠伤寒沙门菌生物被膜形成、氧化应激和毒力作用,为揭示sRNA GcvB在ST生物被膜、氧化应激和毒力中的调控机制提供了研究基础。Small RNAs(sRNAs)are a kind of regulatory molecules that can modulate gene expression at the post-transcriptional level,thereby after the physiological characteristics of bacteria.To investigate the regulatory role of the sRNA GcvB in the virulence of Salmonella typhimurium,in this study,we constructed mutant,complementary and tracer strains of GcvB gene to detect the effects on the ability of biofilm formation and swimming of Salmonella typhimurium.We also analyzed the differences in virulence and infection kinetics between parental strains and deletion strains,predicted and analyzed the potential target genes regulated by GcvB,and explored the role of GcvB in the regulation of the biofilm,oxidative stress and virulence of ST.The results showed that compared with the parental strain and the complementary strain,GcvB deletion increased the biofilm formation ability,significantly enhanced the resistance to oxidative stress,and increased the LD50 to 3.98-fold.Quantitative real-time PCR(qRT-PCR)analysis of the target genes,gcvA,bapA,hilA,igaA and sitD,showed that the expression of gcvA,bapA,igaA,and sitD was up-regulated in the transcriptional level,while the expression of hilA was decreased.The present study confirmed the involvement of sRNA GcvB in Salmonella typhimurium biofilm formation,oxidative stress and virulence,and provided a basis for preliminary studies on the regulatory mechanism of sRNA GcvB in ST biofilm,oxidative stress and virulence.

关 键 词：鼠伤寒沙门菌 GcvB基因 生物被膜 毒力作用 

分 类 号：S852.61[农业科学—基础兽医学]