RNA-Seq分析baeSR和acrB对鼠伤寒沙门菌相关毒力基因表达的影响  

RNA-Seq Analysis of the Effects of baeSR and acrB on the Expression of Virulence Genes Associated with Salmonella typhimurium

作　　者：李锐 高海侠 徐军 王瑞[1] 王文静 张瑞良 赵霞[1] 李琳[1] LI Rui;GAO Haixia;XU Jun;WANG Rui;WANG Wenjing;ZHANG Ruiliang;ZHAO Xia;LI Lin(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China)

机构地区：[1]安徽农业大学动物科技学院

出　　处：《畜牧兽医学报》2019年第11期2318-2325,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31772802);安徽高校自然科学重点研究项目(KJ2017A132)

摘　　要：本研究旨在探索baeSR和acrB基因缺失后对鼠伤寒沙门菌相关毒力基因表达的影响。采用RNA-Seq技术筛选鼠伤寒沙门菌体外诱导环丙沙星耐药株(CR)、acrB基因缺失株(CRΔacrB)、baeSR和acrB基因双缺失株(CRΔbaeSRΔacrB)的差异表达基因,对筛选的基因进行GO富集和KEGG通路富集分析,进一步探究差异表达基因的主要功能及涉及的生物过程,并选出7个差异显著基因进行荧光定量PCR验证。结果表明CRΔacrB与CR相比较,共筛选到1 320个差异表达基因(fold change的绝对值>2,Q-value<0.005),其中上调426个,下调894个;CRΔbaeSRΔacrB与CR相比较,共筛选到1 377个差异表达基因(fold change的绝对值≥2,Q-value<0.005),其中上调405个,下调972个,两个比较组筛选到的毒力相关基因均主要富集在双组分系统、鞭毛组件、沙门菌感染和上皮细胞的细菌入侵等通路中。荧光定量PCR验证结果与RNA-Seq结果基本一致。本研究对baeSR和acrB基因在鼠伤寒沙门菌毒力中的作用进行了初步分析和探索,为进一步研究鼠伤寒沙门菌致病机制奠定基础。The aim of this study was to investigate the effects of baeSR and acrB gene deletions on the expression of virulence genes associated with Salmonella typhimurium. The differentially expressed genes of ciprofloxacin-resistant strain(CR), acrB gene-deficient strain(CRΔacrB), baeSR and acrB gene double-deletion strain(CRΔbaeSRΔacrB) were screened by using RNA-Seq technique. GO enrichment and KEGG pathway enrichment analysis were carried out on the selected genes to further explore the main functions of differentially expressed genes and the biological processes involved, and 7 genes with significant differences were selected for verification by real-time PCR. The results showed that, a total of 1 320 differentially expressed genes(absolute value of Fold change>2, Q-value<0.005) were screened between CRΔacrB and CR, of which 426 were up-regulated and 894 were down-regulated. There were 1 377 differentially expressed genes(absolute value of Fold change ≥2, Q-value<0.005) were screened between CRΔbaeSRΔacrB and CR, of which 405 were up-regulated and 972 were down-regulated. The virulence-related genes screened by the two comparison groups were mainly enriched in pathways such as two-component system, flagellar component, Salmonella infection, and bacterial invasion of epithelial cells. The results of real-time PCR are basically consistent with those of RNA-Seq. This study preliminarily analyzed and explored the role of baeSR and acrB genes in the virulence of Salmonella typhimurium, laying a foundation for further study on the pathogenic mechanism of Salmonella typhimurium.

关 键 词：鼠伤寒沙门菌 RNA-SEQ baeSR ACRB 荧光定量PCR 

分 类 号：R378.22[医药卫生—病原生物学]