鼠伤寒沙门菌baeSR和acrB双基因缺失株相关耐药基因的RNA-seq分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0464

摘要/Abstract

摘要： 应用转录组测序技术（RNA-seq）筛选鼠伤寒沙门菌baeSR和acrB相关的靶基因,为鼠伤寒沙门菌耐药机制的深入研究奠定基础。采用RNA-seq技术分析baeSR和acrB基因缺失后表达量变化的相关耐药基因,通过GO富集和KEGG通路探索其主要功能,qRT-PCR验证部分差异基因。以 |log₂（Fold Change）| > 1且q value < 0.005为条件,其中以CR△acrB和CR为比较组,共筛选到1 320个差异表达基因,894个下调,426个上调;以CR△baeSR△acrB和CR为比较组,共筛选到1 377个差异表达基因,972个下调,405个上调。GO 富集分类结果显示差异基因主要集中在跨膜运输、细胞黏附、纤毛或鞭毛运动等功能;KEGG 数据库显示,差异基因主要富集在代谢途径、ABC转运系统、双组份信号转导系统、鞭毛组装、β-内酰胺抗性等通路。其中,ompR、csgD、fliA、fliG可通过调节细菌生物膜形成能力调控其对多种抗生素的耐药性;emrA、mdtC、yejE属于外排泵相关基因,可介导细菌对黏菌素、新生霉素等的耐药性;STM3031可赋予沙门菌头孢曲松抗性;pmrF可参与调节细菌对多粘菌素的抗性。鼠伤寒沙门菌acrB和baeSR基因缺失可影响相关耐药基因的表达。

关键词: 鼠伤寒沙门菌, baeSR, acrB, RNA-Seq, 相关耐药基因

Abstract: The transcriptome sequencing technology（RNA-seq）was used to screen the target genes related to baeSR and acrB of Salmonella typhimurium,which laid a foundation for further study on the drug resistance mechanism of S. typhimurium. RNA-Seq was applied to screen the related drug-resistant genes with varied expressions after the deletion of baeSR and acrB genes,GO enrichment and KEGG pathways to explore their functions,and qRT-PCR to verify some differential genes. Under the condition of |log₂（Fold Change）| > 1 and q value < 0.005,a total 1 320 differentially expressed genes were screened by CR△acrB/CR as comparison group,of which 894 were down-regulated and 426 were up-regulated. A total 1 377 differentially expressed genes were screened by CR△baeSR△acrB/CR,of which 972 genes were down-regulated and 405 genes were up-regulated. GO enrichment classification results showed that the differential genes were mainly concentrated in transmembrane transport,cell adhesion,cilia or flagellum movement and other functions. KEGG database showed that the differential genes were mainly concentrated in metabolic pathways,ABC transport system,two-component signal transduction system,flagellum assembly,beta-lactam resistance and so on. Among them,ompR,csgD,fliA and fliG regulated their resistances to various antibiotics by regulating the ability of bacterial biofilm formation. emrA,mdtC and yejE belonged to efflux pump-related genes,which mediated the resistance of bacteria to colistin and neomycin. STM3031 endowed Salmonella with ceftriaxone resistance,and pmrF regulated bacterial resistance to polymyxins. In conclusion,the deletion of acrB and baeSR genes in S. typhimurium may affect the expressions of related resistance genes.

Key words: Salmonella typhimurium, baeSR, acrB, RNA-Seq, related drug-resistant genes

徐军, 王瑞, 李锐, 高海侠, 张瑞良, 王文静, 赵霞, 李琳. 鼠伤寒沙门菌baeSR和acrB双基因缺失株相关耐药基因的RNA-seq分析[J]. 生物技术通报, 2019, 35(12): 76-84.

XU Jun, WANG Rui, LI Rui, GAO Hai-xia, ZHANG Rui-liang, WANG Wen-jing, ZHAO Xia, LI Lin. RNA-seq Analysis of Drug-resistant Genes Associated with baeSR and acrB Double Gene Deletion Strains of Salmonella typhimurium[J]. Biotechnology Bulletin, 2019, 35(12): 76-84.

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