牛乳源金黄色葡萄球菌EsxB蛋白的表达及免疫效应分析  

Expression and Analysis of Immunogenicity of EsxB Protein of Staphylococcus aureus Isolated from Bovine Milk

作　　者：张雪静 易鸳鸯 杨志荣 马晓慧 苏艳[1] ZHANG Xue-jing;YI Yuan-yang;YANG Zhi-rong;MA Xiao-hui;SU Yan(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业大学动物医学学院

出　　处：《新疆农业大学学报》2019年第1期53-58,共6页Journal of Xinjiang Agricultural University

基　　金：新疆维吾尔自治区高校科研计划项目(XJEDU2018I009)

摘　　要：为分析牛乳源金黄色葡萄球菌(Staphylococcus aureus)EsxB蛋白的免疫特性,扩增EsxB基因并构建了pET-28a-EsxB重组质粒,转化后的重组菌经诱导表达后用SDS-PAGE和Western blotting检测重组蛋白;以纯化的EsxB蛋白免疫小鼠,检测其抗体水平、滴度及亚型、测定攻击保护率和观察小鼠肺、肾病理组织学变化。结果显示,成功纯化了EsxB蛋白,且该蛋白具有良好的抗体反应特性。免疫小鼠的血清抗体水平与对照组相比显著升高;抗体滴度可达1∶24 300,抗体亚型以IgG1和IgG2b为主,免疫小鼠的攻击保护率可达75%;且免疫后降低了S.aureus攻击对小鼠肺脏及肾脏的病理损伤。重组EsxB蛋白具有良好的免疫原性和保护效果。In order to analyze the immunological characteristics of EsxB protein of Staphylococcus aureus isolated from dairy milk,the EsxB gene was amplified and EsxB-pET-28a recombinant plasmid was constructed.The expression and purified EsxB was analyzed by SDS-PAGE and Western blotting.The purified EsxB was used to immunize mice to check the antibody level,titer and subtybe in serum of immunized mice.Then the challenge protection and histopathological examination were investigated after challenge with S.aureus.The result indicated that the EsxB was successfully purified and the protein had better antibody reaction;The EsxB induced high levels of specific IgG antibody in serum,the antibody titer could reach 1∶23 400,the antibody subtype mainly were IgG1,IgG2b.The challenge-protection rate of EsxB was 75%and EsxB immunization could significantly reduce the histopathology damage of lung and kidney in mice.In conclusion,the recombinant EsxB protein had better immunogenicity and immunoprotective capacity.

关 键 词：牛源金黄色葡萄球菌 EsxB蛋白 表达与纯化 免疫效应 

分 类 号：S852.61[农业科学—基础兽医学]