亚洲带绦虫包虫诊断抗原P-29基因克隆表达  

Cloning,prokaryotic expression and immunoreacticity of hydatid disease diagnostic antigen P-29 of Taenia saginata asiatica

作　　者：周灵贵[1] 戴佳琳[1] 黄江[1] 廖兴江[1] 胡旭初[2] 余新炳[2] 申萍香[1] 郎书源[1] 

机构地区：[1]贵阳医学院多媒体形态学实验室,贵阳550004 [2]中山大学中山医学院人体寄生虫教研室

出　　处：《中国公共卫生》2009年第9期1090-1092,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金(30760227);贵州省科技攻关项目(2008-3060)

摘　　要：目的识别亚洲带绦虫包虫诊断抗原P-29(Hydatid disease diagnostic antigen P-29)的已知序列并行克隆和蛋白表达及免疫学研究。方法利用在线生物信息学工具序列分析后以亚洲带绦虫成虫包虫诊断抗原P-29基因克隆到原核表达质粒pET-28a(+)中,在大肠埃希菌BL21/DE3中诱导表达,重组后的蛋白用His-镍蛋白纯化柱纯化,纯化的重组蛋白用蛋白印迹进行免疫学分析。结果重组体构建成功并以包涵体形式存在,破包涵体纯化蛋白并得到高纯度的蛋白,且该重组蛋白可被亚洲带绦虫及牛带绦虫病人血清识别,表明其具有免疫反应性。结论亚洲带绦虫成虫包虫诊断抗原P-29可在原核表达系统中获得具有免疫活性的高效表达。Objective To identify the structure and function of hydatid disease diagnostic antigen P-29 of Taenia saginata asitica, and to construct prokaryotic recombinant plasmids of the gene, to express and purify the recombinant protein and conduct a preliminary immunoreacticity study. Methods By online gene analysis at bioinfomatics websites, the gene from the Taenia saginata asiatica full-length cDNA plasmid libratory was identified. The coding region sequence and the characteristics of the deduced protein were analyzed. The recombinant protein was detected by SDS-PAGE after being in- duced with IPTG in E coli BL21/DE3 and purified with NI-IDA affinity chromatography. Its immunoreactivity was determined by Western blotting. Results The recombinant plasmid was successfully constructed. The recombinant protein could react with Taenia asiatica and Taeniarhynchus saginatus infected patient＇s serum. Conclusion The hydatid disease diagnostic antigen P-29 of Taenia saginata asitica was cloned and expressed, and the purified protein was confirmed with immunogenicity.

关 键 词：亚洲带绦虫 包虫诊断抗原P-29 基因克隆 原核表达 免疫反应性 

分 类 号：R383.32[医药卫生—医学寄生虫学]