牛结核分枝杆菌抗原蛋白Ag85B的表达及纳米疫苗的制备  

Expression of Mycobacterium Bovis Antigen Protein Ag85B and Preparation of Its Nano-vaccine

作　　者：杜军[1] 邓光存[1] 

机构地区：[1]宁夏大学生命科学学院,银川750021

出　　处：《湖北农业科学》2015年第6期1421-1424,1429,共5页Hubei Agricultural Sciences

基　　金：宁夏自然科学基金项目(NZ13036)

摘　　要：以牛结核分枝杆菌(Mycobacterium bovis,MTB)C68001株基因组DNA为模板,克隆免疫优势抗原基因Ag85B,构建重组表达质粒p ET-28a-ag85b,转化入大肠埃希菌BL21(DE3),经IPTG诱导表达重组蛋白,对表达产物进行SDS-PAGE和Western blot分析。以明胶为佐剂,Ag85B为目的抗原制备纳米疫苗,并对其进行了相关的工艺检测。以BCA法建立测定蛋白质含量的标准曲线,在此基础上检测纳米疫苗的载药率和包封率。结果显示,明胶佐剂疫苗的载药率为71.83%,包封率为41.03%,符合纳米疫苗的工艺要求。The immuno-dominant antigen gene Ag85B was cloned using the genomic DNA of Mycobacterium bovis strain C68001 as template and used to construct the recombinant plasmid pET-28a-ag85b, transformed into E. coli BL21 （DE3）. Induced by IFFG, the expressed recombinant protein was analyzed by SDS-PAGE and Western blot method. The nano-vaccine was prepared using gelatin as adjuvant and Ag85B as objective antigen. The related technology was tested. Based on the protein standard curve obtained with BCA method. The drug loading and encapsulation efficiency of the nano-vaccine was 71.83% and 41.03%, respectively, metting the technological requirements of nano-vaccine.

关 键 词：牛结核分枝杆菌(Mycobacterium bovis MTB) Ag85B抗原 原核表达 纳米疫苗 

分 类 号：R392.11[医药卫生—免疫学]