摘要
[目的]筛选可结合Caco-2细胞表面蛋白的副溶血弧菌外膜蛋白。[方法]筛选副溶血弧菌外膜中的黏附蛋白,将Caco-2细胞表面蛋白生物素化并固定于中性卵白素树脂上,进一步利用亲和层析技术来筛选副溶血弧菌的外膜蛋白。[结果]通过LC-MS/MS质谱技术鉴定出3个候选蛋白:ATP synthase subunit alpha、ATP synthase subunit beta和outer membrane protein U。通过对这3个蛋白进行克隆基因和原核表达,成功纯化得到相应重组蛋白。通过进一步的间接免疫荧光试验,发现3种蛋白对于Caco-2细胞均有黏附作用。[结论]推测ATP synthase subunit alpha、ATP synthase subunit beta和outer membrane protein U这3种蛋白可能是潜在的黏附因子。
[Objective]To screen the outer membrane protein of Vibrio parahaemolyticus(VP)binding to Caco-2 cell surface proteins.[Method]In order to screen adhesins of VP,the cell surface proteins of Caco-2 cells were biotinylated and then employed as the baits to capture host binding outer membrane proteins of VP by using affinity chromatography.[Result]ATP synthase subunit alpha,ATP synthase subunit beta and outer membrane protein U were identified by LC-MS/MS approach.The three recombinant candidate proteins were expressed and purified,furtherly their binding activity to Caco-2 cells were evaluated via indirect immune-flourescence assays.[Conclusion]The three outer membrane proteins of VP which are capable of binding to Caco-2 cell surface proteins are candidates of VP adhesion.
作者
陈萌
李卓昱
鲍玉杰
袁增智
CHEN Meng;LI Zhuo-yu;BAO Yu-jie(College of Life Sciences,Tianjin Normal University,Tianjin 300387)
出处
《安徽农业科学》
CAS
2018年第12期118-121,共4页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金项目(31402322)
天津市应用基础与前沿技术研究计划项目(15JCQNJC46400)
天津师范大学博士基金项目(52XB1412)
关键词
副溶血弧菌
外膜蛋白
黏附因子
Vibrio parahaemolyticus
Outer membrane protein
Adhesin
作者简介
陈萌(1992—),女,天津人,硕士研究生,研究方向:水生生物分子生物学。;通讯作者,袁增智,讲师,博士,从事水生动物传染病学研究。
