肠炎沙门菌greB基因缺失株构建及其生物学特性初步研究  

Construction and biological characterization of greB gene deletion strain of Salmonella Enteritidis

作　　者：蔡梦雷 姜婷婷 吴村 刘东海 焦晓丽 张凯迪 荣丽娟 王俊 崔国林 CAI Meng-lei;JIANG Ting-ting;WU Cun;LIU Dong-hai;JIAO Xiao-li;ZHANG Kai-di;RONG Li-juan;WANG Jun;CUI Guo-lin(Hebei University of Engineering,Handan 056038,China;Jiaxing Vocational and Technical College,Jiaxing 314036,China)

机构地区：[1]河北工程大学,邯郸056038 [2]嘉兴职业技术学院,嘉兴314036

出　　处：《中国人兽共患病学报》2023年第4期325-332,共8页Chinese Journal of Zoonoses

基　　金：河北省自然科学基金项目(No.C2019402120);河北省高等学校科学技术研究项目(No.QN2019015);嘉兴职业技术学院人才启动基金专项(No.20407030086)。

摘　　要：目的研究旨在探究转录延长因子GreB对肠炎沙门菌致病力相关生物表型的影响。方法以肠炎沙门菌ATCC13076为亲本株,利用λ-Red重组系统构建ΔgreB基因缺失株及其回补株,测定肠炎沙门菌及其衍生菌在多种抑菌环境中生存能力、对细胞感染能力及在雏鸡体内繁殖动态,并采用RNA-seq检测和分析GreB对肠炎沙门菌致病力相关基因转录的影响。结果结果显示,greB基因缺失对肠炎沙门菌体外生长、生物被膜形成、SDS和高渗透压抵抗力无显著影响,但ΔgreB基因缺失株抗氧化能力显著提升;greB基因缺失减弱肠炎沙门菌对HCT116细胞入侵;与亲本株相比,ΔgreB基因缺失株在雏鸡肝脏、脾脏和肺脏的载量均下降,对雏鸡致病力下降约4倍;greB基因缺失导致肠炎沙门菌85个基因差异表达,其中34个基因表达下调,主要包括fim操纵子和inv操纵子,51个基因表达上调,主要包括pdu操纵子和ssa操纵子。结论greB基因缺失引致肠炎沙门菌多种毒力相关基因差异转录表达,增强肠炎沙门菌抗氧化能力,但降低其对上皮细胞侵袭力和动物致病力。The aim of this study was to investigate the effects of the transcription elongation factor GreB on the virulence-related phenotypes of Salmonella Enteritidis.Salmonella Enteritidis ATCC13076 was used as the parent strain,and anΔgreB gene deleted strain and its trans-complemented strain were constructed with theλ-Red recombination system.The viability of Salmonella Enteritidis and its derivatives in a variety of bacteriostatic environments,the ability to infect cells and the reproduction dynamics in chickens were detected with the plate counting method.The effects of GreB on the transcription of virulence-related genes of Salmonella Enteritidis was detected with RNA-seq.The results indicated that greB gene deletion had no significant effects on growth,biofilm formation,SDS and high osmotic pressure resistance of Salmonella Enteritidis in vitro,but the antioxidant capacity of theΔgreB gene deleted strain was significantly greater than that of the parent stain.The greB gene deletion attenuated the ability of Salmonella Enteritidis to enter HCT116 cells.TheΔgreB gene deleted strain had a lower load in the liver,spleen and lungs of chicks,and approximately four times lower virulence to chicks than the parent strain.After the greB gene was deleted,85 genes were differentially expressed,among which 34 genes were down-regulated,mainly including the fim operon and inv operon,whereas 51 genes were up-regulated,mainly including the pdu operon and ssa operon.These results suggest that greB gene deletion leads to differential transcription of virulence-related genes of Salmonella Enteritidis,and enhances its oxidation resistance but decreases its invasiveness to epithelial cells and its virulence to animals.

关 键 词：肠炎沙门菌 greB基因 λ-Red重组系统 生物学特性 RNA-seq 

分 类 号：S852.61[农业科学—基础兽医学] R378.2[农业科学—兽医学]