基于T3SS的沙门氏菌抑制剂筛选系统构建与多酚类化合物活性评价OA
Construction of a Screening System for Salmonella Inhibitors Based on T3SS and Activity Evaluation of Polyphenolic Compounds
Ⅲ型分泌系统(type Ⅲ secretion system,T3SS)是沙门氏菌重要毒力因子,针对T3SS的特异性抑制剂是抗沙门氏菌感染的有效方法.用Red同源重组技术构建鼠伤寒沙门氏菌invA基因缺失株(SL1344ΔinvA),用无缝克隆连接构建pWSK29-sopE2-luc报告质粒,分别转化SL1344和SL1344ΔinvA获得报告菌株,以MOI=100感染复数建立鼠伤寒沙门氏菌感染HeLa细胞模型,建立T3SS抑制剂筛选系统,并筛选多酚类化合物以获得T3SS天然产物抑制剂.结果显示,成功获得SL1344ΔinvA菌株及SL1344(pWSK29-sopE2-luc)和 SL1344ΔinvA(pWSK29-sopE2-luc)报告菌株,通过 T3SS 抑制剂筛选系统,筛选获得没食子酸乙酯、没食子酸丙酯、咖啡酸、4-香豆酸和单宁酸T3SS有效抑制剂.本研究成功构建T3SS抑制剂筛选系统,为沙门氏菌抗毒力因子的药物筛选搭建平台.
The type Ⅲ secretion system(T3SS)is a crucial virulence factor of Salmonella.Specifie inhibi-tors targeting T3SS is considered an effective method for combating Salmonella infections.This study uti-lized Red homologous recombination technology to construct an invA gene deletion strain of Salmonella Ty phi murium(SL1344 ΔinvA).A seamless cloning method was employed to construct the pWSK29-sopE2-luc reporter plasmid.HeLa cell infection model of Salmonella typhimurium was established at a multiplicity of infection(MOI)of 100 to set up a screening system for T3SS inhibitors.Polyphenolic com-pounds were screened to obtain natural product inhibitors of T3SS.The results show that the SL1344ΔinvA strain,as well as the SL1344(pWSK29-sopE2-luc)and SL1344ΔinvA(pWSK29-sopE2-luc)reporter strains,were successfully obtained.Through the T3SS inhibitor screening system,this study suc-cessfully identified effective T3SS inhibitors,including ethyl gallate,propyl gallate,caffeic acid,p-coumaric acid and tannic acid.This study successfully established a screening system for T3SS inhibitors,providing a platform for drug screening targeting virulence factors of Salmonella,and evaluated polyphenolic com-pounds to identify effective T3SS inhibitors.
茹梦珂;刘盼盼;李穗湘;杨珍;郝宝成;王胜义;胡庭俊;崔东安
广西大学动物科学技术学院,广西南宁 530005||中国农业科学院兰州畜牧与兽药研究所,甘肃兰州 730050兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室,甘肃兰州 730000||中国农业科学院兰州畜牧与兽药研究所,甘肃兰州 730050广西大学动物科学技术学院,广西南宁 530005||中国农业科学院兰州畜牧与兽药研究所,甘肃兰州 730050中国农业科学院兰州畜牧与兽药研究所,甘肃兰州 730050中国农业科学院兰州畜牧与兽药研究所,甘肃兰州 730050中国农业科学院兰州畜牧与兽药研究所,甘肃兰州 730050广西大学动物科学技术学院,广西南宁 530005兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室,甘肃兰州 730000||中国农业科学院兰州畜牧与兽药研究所,甘肃兰州 730050
农业科技
鼠伤寒沙门氏菌Ⅲ型分泌系统(T3SS)毒力抑制Red同源重组多酚类化合物
Salmonella typhimuriumType Ⅲ secretion system(T3SS)Virulence inhibitionRed homolo-gous recombinationPolyphenols
《动物医学进展》 2026 (3)
1-8,8
国家重点研发计划项目(2023YFD1800802)
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