荧光假单胞菌SlyB的原核表达、抗原性分析及抗血浆杀菌作用  

Prokaryotic expression,antigenicity analysis and anti-plasma bactericidal effect of Pseudomonas fluorescens outer membrane lipoprotein SlyB

作　　者：孙薇 荣娜 简思杰 晁嘉 刘祥[1,2] 丁锐[1] 陈锐[1] SUN Wei;RONG Na;JIAN Si-jie;CHAO Jia;LIU Xiang;DING Rui;CHEN Rui(School of Bioscience and Engineering,Shaanxi University of Technology,Hanzhong 723000,China;Chinese-German Joint Institute for Naturel Product Research,Shaanxi University of Technology,Hanzhong 723000,China)

机构地区：[1]陕西理工大学生物科学与工程学院,陕西汉中723000 [2]陕西理工大学中德天然产物研究所,陕西汉中723000

出　　处：《陕西理工大学学报（自然科学版）》2022年第1期49-57,共9页Journal of Shaanxi University of Technology:Natural Science Edition

基　　金：国家外国专家局项目(GDT20186100426);陕西省科技厅重点研发计划项目(2020LSFP2-38);陕西理工大学人才启动项目(SLGQD1803)。

摘　　要：采用生物信息学方法分析SlyB的理化性质,分子克隆表达纯化获得SlyB蛋白并免疫小鼠制备SlyB抗血清,Western blotting检测抗血清特异性与效价,免疫酶联法模拟SlyB抗血清对鱼类主要致病菌的体外免疫识别作用,Western blotting分析SlyB蛋白抵抗鱼血浆的杀菌作用。结果显示:SlyB蛋白在假单胞菌属和杆菌属间同源性较好;获得SlyB最佳诱导表达条件;验证SlyB抗血清特异性较好,效价达到1∶12800;ELISA显示SlyB抗血清对嗜水气单胞菌、荧光假单胞菌和溶藻弧菌存在免疫识别作用,Western blotting表明SlyB蛋白对细菌抵抗鱼血浆的杀菌作用呈下调趋势。表明SlyB蛋白具有较好的抗原性,在蛋白亚单位疫苗上有应用前景。Bioinformatics methods were used to analyze the physical and chemical properties of SlyB.SlyB protein was obtained by molecular cloning,expression and purification,and mice were immunized to prepare SlyB antiserum.Western blotting was used to detect the specificity and titer of the antiserum.In vitro immune recognition of pathogenic bacteria.Western blotting analysis of SlyB protein against the bactericidal effect of fish plasma.The results showed that the SlyB protein had good homology between Pseudomonas and Bacillus,the optimal conditions for inducing expression of SlyB were obtained.The specificity of SlyB antiserum has been verified to be better,with titer reaching 1∶12800,and ELISA has shown that SlyB antiserum has immunorecognition effect on Aeromonas hydrophila,Pseudomonas fluorescens and Vibrio alginolyticus.Western blotting has shown that SlyB protein displays a downward trend in the bactericidal effect of bacteria against fish plasma.It shows that SlyB protein has good antigenicity and has application prospects in protein subunit vaccines.

关 键 词：荧光假单胞菌 外膜脂蛋白SlyB 多克隆抗血清 抗血浆杀菌作用 

分 类 号：Q71[生物学—分子生物学]