牛巴贝斯虫新疆株MSA-2c基因的克隆表达及其重组蛋白免疫原性  

Cloning and expression of merozoite surface antigen 2 locus(MSA-2c) of Babesia bovis isolated in Xinjiang and antigenicity of its recombinant protein

作　　者：简子健[1] 袁江玲[1] 马素贞[1] 黄家雨[1] 沈炯玉[1] 

机构地区：[1]新疆农业大学动物医学院,新疆乌鲁木齐830052

出　　处：《中国兽医学报》2009年第10期1282-1285,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(3066141)

摘　　要：本试验将新疆株牛巴贝斯虫的MSA-2c基因克隆,并构建重组质粒pGEX-4T-2/MSA-2c。利用大肠杆菌原核表达系统进行外源蛋白的表达,并成功诱导出GST-MSA-2c融合蛋白。通过优化诱导条件,得到了较高的可溶性表达;利用亲和层析技术纯化后的重组蛋白皮下免疫试验小鼠。间接ELISA检测发现,免疫接种56 d后,抗重组蛋白的抗体效价达到1∶220000以上。用获得的抗血清进行Western-blot试验,可获得清晰的免疫反应条带。结果表明,本试验所表达的MSA-2c蛋白与文献报道的目的蛋白相符,并具有免疫原性。同时本试验也为建立以MSA-2c重组蛋白作为抗原的血清学诊断体系奠定了基础。The merozoite surface antigen 2 locus gene （MSA-2c） of Babesia boris found in Xinjiang was cloned and ligated into pGEX-4T-2 to construct a recombinant plasmid,pGEX-4T-2/MSA-2c. A fusion protein,GST-MSA- 2c,was successfully expressed through prokaryotic expression system in E. coli. The high quantity of soluble expression was obtained by improving the induction conditions. The recombinant protein purified by affinity chromatography was inoculated into mice subcutaneously. The antibody titers of the recombinant protein in murine sera were above 1 ： 220 000 at 56 days postinoculation by indirect ELISA. The antisera were then examined by Western-blot and a clear reaction strip was found. The results showed that of the expressed MSA-2c protein was identified with interest protein reported in records,and was highly immunogenic （antigenic）. It laysa foundation of construction serologically diagnostic system using MSA-2c recombinant protein as a diagnostic antigen.

关 键 词：牛巴贝斯虫 MSA-2c 重组表达 免疫原性 

分 类 号：S852.72[农业科学—基础兽医学]