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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 known sequence parallel cloning and protein expression and immunological studies of the hydatid disease diagnostic antigen P-29. Methods The sequence analysis of online bioinformatics tools was carried out. The diagnostic antigen P-29 of the Asian female Taenia asiatis was cloned into prokaryotic expression plasmid pET-28a (+) and induced in Escherichia coli BL21 / DE3. After recombination Purified recombinant protein was subjected to immunological analysis by Western blotting. Results The recombinant protein was successfully constructed and existed as inclusion body. The inclusion body was purified and the protein was purified. The purified recombinant protein was identified by the serum of Taenia asiatis and Taenia saginata as immunoreactive. Conclusion The diagnostic antigen P-29 of the adult worm hydatidstate can be highly expressed in prokaryotic expression system.