摘要
为挖掘介导肠炎沙门菌运动能力的功能基因,深入研究沙门菌运动及致病机制,采用转座子突变技术构建肠炎沙门菌C50041突变库,筛选获得运动性弱或无的突变株;在突变株运动缺陷表型确认后,提取细菌基因组DNA,PCR扩增转座子在细菌基因组中所插入的侧翼序列,并对侧翼序列进行基因测序进行Blast比对相似度,确定突变失活的基因。结果显示:从Tn5转座子突变库中筛选了10000多株突变株,从中筛选出10株运动性缺陷的突变株,并成功PCR扩增每个突变株转座子侧翼序列,通过Blast比对初步确定fliP、fliD、rfaL、rfbK、cpsG、csrD 6个基因的失能导致肠炎沙门菌运动缺陷,其中fliD、fliP和cpsG为鞭毛合成相关基因,rfbK和rfaL为脂多糖合成相关基因,csrD为调控基因。研究结果为进一步探讨沙门菌运动性及致病机制奠定了基础。
In order to mine genes mediating bacterial motility for further study on mechanism of Salmonella motility and pathogenicity,Tn5 transposon was used to generate a library of mutants within the Salmonella Enteritidis genome,and mutants with weak or no motility would be screened out.After defect phenotype being confirmed,bacterial genomic DNA was extracted,the sequence flanking with Tn5 transposon was amplified by PCR and sequenced,then compared by Blast alignment,the gene inserted by transposon was determined by its identity.The results showed that 10 motility-deficient mutants were screened out from over10 000 Tn5 transposon mutants,and 6 genes including fliP,fliD,rfaL,rfbK,cpsG,csrD were determined.Among them,fliD and fliP were flagellar synthesis-related genes,rfaL,rfbK and cpsG were lipopolysaccharide synthesis-related genes and csrD was regulatory gene.These results would provide a base for further study of these genes involved in the motility and pathogenicity of Salmonella Enteritidis.
作者
黄鹏
陈敬
蔡媛
刘桂凤
耿士忠
HUANG Peng;CHEN Jing;CAI Yuan;LIU Guifeng;GENG Shizhong(Key Laboratory of Zoonoses of Jiangsu Province,Y angzhou University,Y angzhou,Jiangsu 225009;Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses of Jiangsu Province,Y angzhou,Jiangsu 225009;Key Laboratory of Prevention and Control of Biological Hazard Factors(Animal Origin)for Agifood Safety and Quality,Yangzhou University,Yangzhou,Jiangsu 225009;Joint International Research Laboratory of Agriculture&Agri-Product Safety,Ministry of Education of China,Yangzhou Universily,Yangzhou,Jiangsu 225009)
出处
《中国家禽》
北大核心
2022年第3期44-49,共6页
China Poultry
基金
国家重点研发项目(2016YFD0501607)
江苏省自然科学基金项目(BK20151306)
江苏省重点实验室开放课题项目(0273896034819)。
关键词
肠炎沙门菌
运动性
转座子
鞭毛
功能基因
SalmonellaEnteritidis
motility
transposon
flagellar
functional genes
