重组牛乳源金黄色葡萄球菌GapC蛋白优势B细胞抗原表位的预测和筛选  

Prokaryotic Expression of Recombinant Bovine Mastitis Staphylococcus aureus GapC Protein and Identification of Its B-cell Epitopes

作　　者：陈晓萌 张雪静 张欢 张宝江[1] 苏艳[1] CHEN Xiao-meng;ZHANG Xue-jing;ZHANG Huan;ZHANG Bao-jiang;SU Yan(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052)

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《生物技术通报》2023年第5期306-313,共8页Biotechnology Bulletin

基　　金：新疆维吾尔自治区高校科研计划重点项目(XJEDU2018I009)。

摘　　要：旨在表达牛乳源金黄色葡萄球菌(Staphylococcus aureus)GapC蛋白并对其B细胞抗原表位进行预测与鉴定,本研究利用实验室分离鉴定的S.aureus分离株15119扩增GapC基因并构建重组表达质粒pET-28a-GapC,诱导纯化得到分子量为44 kD重组蛋白GapC,以此免疫新西兰大白兔,获得特异多克隆抗体。利用生物信息学方法,对GapC蛋白的二级及三级结构进行分析,预测其B细胞抗原表位,并利用特异性抗体对筛选的表位进行鉴定。结果表明,GapC蛋白具有良好的免疫原性,筛选出7个线性B细胞抗原表位,利用兔抗重组GapC蛋白多克隆抗体鉴定得到了PL 5(^(221)IPEIDGKLDGGAQRVP^(236))多肽和PL 7(^(264)KNASNESFGYTEDEIVSSDVVGM^(286))2个优势B细胞表位。本研究成功制备了GapC蛋白,预测并鉴定了2个优势抗原表位,为其嵌合表位疫苗的开发提供了技术支持。To express the GapC protein of Staphylococcus aureus isolated from bovine milk and to predict and identify its B-cell epitope,GapC gene of S.aureus strain 15119 isolated by our lab was amplified and the recombinant expression plasmid pET-28a-GapC was constructed.The recombinant protein GapC with a molecular weight of 44 kD was induced and purified.Then,the purified protein was used to immunize New Zealand white rabbits to obtain polyclonal antibodies.And bioinformatics method was to analyze the secondary and tertiary structures of GapC protein and predict the B cell epitopes.The predicted and selected epitopes were identified by specific antibodies.The results showed that the GapC protein had promising immugenicity and 7 B cell epitopes were selected.Two of dominant B cell epitope PL5(^(221)IPEIDGKLDGGAQRVP^(236))and PL7(^(264)KNASNESFGYTEDEIVSSDVVGM^(286))were identified using specific polyclonal antibody.This data lays the foundation for the development of epitope vaccine.

关 键 词：牛源金黄色葡萄球菌 GapC蛋白 表达 B细胞抗原表位 预测与鉴定 

分 类 号：R73[医药卫生—肿瘤]