GreA蛋白对肠炎沙门菌生物学特性及致病力影响  

The Effect of GreA Protein on the Biological Characteristics and Pathogenicity of Salmonella Enteritidis

作　　者：蔡梦雷 赵东旭 张政钢 刘东海 姜婷婷 苏士炫 闫雪敏 薛晓阳 崔国林 CAI Menglei;ZHAO Dongxu;ZHANG Zhenggang;LIU Donghai;JIANG Tingting;SU Shixuan;YAN Xuemin;XUE Xiaoyang;CUI Guolin(School of Life Science and Food Engineer,Hebei University of Engineering,Handan 056038,China;Huayu Agricultural Science and Technology Co.,Ltd,Handan 057151,China)

机构地区：[1]河北工程大学生命科学与食品工程学院,邯郸056038 [2]华裕农业科技有限公司,邯郸057151

出　　处：《畜牧兽医学报》2024年第11期5173-5182,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：河北省自然科学基金项目(C2019402120);河北省高等学校科学技术研究项目(QN2019015);河北省三期现代农业产业技术体系蛋禽创新团队资助项目(HBCT2024260205)。

摘　　要：旨在研究转录延长因子GreA蛋白对肠炎沙门菌生物学特性和致病力的影响。以ATCC13076为亲本株,通过λ-Red重组系统构建ΔgreA和ΔgreB单基因缺失株、ΔgreAΔgreB双基因缺失株,检测亲本株及缺失株的体外生长和运动力,对压力环境抵抗力,生物被膜形成能力,以及对细胞和小鼠的致病力,并利用RNA-seq检测差异表达基因。结果显示:GreA蛋白负调控肠炎沙门菌H2O2抵抗力、生长速度以及生物被膜形成能力,正调控肠炎沙门菌运动能力、细胞入侵和胞内增殖能力及其在小鼠脏器的定殖能力,但对高温和酸性环境抵抗力无显著影响。GreA蛋白影响107个基因表达,其中正向调控fim操纵子、flg操纵子、inv操纵子等,主要与细菌运动及入侵细胞相关;负向调控于cps操纵子、cys操纵子、leu操纵子等,主要与氨基酸合成及代谢相关。GreA蛋白通过调控鞭毛合成和细胞入侵相关代谢通路的表达,影响肠炎沙门菌环境适应性生存及致病力。The aim is to study the effect of GreA protein on the biological characteristics and pathogenicity of Salmonella Enteritidis.Based on the parent strain ATCC13076,ΔgreA andΔgreB single gene deletion strains andΔgreAΔgreB double gene deletion strains were constructed byλ-Red recombinant system.Their growth and swimming motility,resistance to stressful environment,biofilm formation ability,and pathogenicity to cells and mice of the parents and the deletion strains were detected.RNA-seq was used to detect differentially expressed genes(DEGs).GreA protein negatively regulated the H 2O 2 resistance,the growth in vitro and biofilm formation ability of Salmonella Enteritidis,and positively regulated the motor ability,cell invasion and intracellular proliferation ability of Salmonella Enteritidis and its colonizing ability in mouse organs,but have no significant effect on resistance of Salmonella Enteritidis to high temperature and acid environment.GreA protein affects the expression of 107 genes,including positive regulation of inv operon,fim operon and flg operon,which are mainly related to bacterial motility and cell invasion;and negative regulation of cps operon,cbp operon and leu operon,which are related to amino acid synthesis and metabolism.GreA protein affects the environmental adaptability and pathogenicity of Salmonella Enteritidis by regulating the expression of metabolic pathways related to flagellar synthesis and cell invasion.

关 键 词：肠炎沙门菌 GreA 生物学特性 RNA-SEQ 

分 类 号：S852.612[农业科学—基础兽医学]