保定地区奶牛微小隐孢子虫Cp23基因的原核表达及免疫原性分析

Prokaryotic expression and immunogenicity analysis of Cp23 gene of Cryptosporidium parvum in Baoding area

采用PCR技术扩增了微小隐孢子虫Cp23基因,将Cp23基因克隆至原核表达载体pET-28a（＋）中,构建重组表达载体pET28a-Cp23。将其转化至宿主菌E.coli BL21（DE3）中,用IPTG诱导表达后进行SDS-PAGE和Western blot分析。SDS-PAGE结果表明,蛋白相对分子质量为25 000,目的蛋白高效表达,且为包涵体蛋白。Western blot结果表明,表达产物可被微小隐孢子虫阳性血清识别,具有良好的免疫反应原性。本研究为抗微小隐孢子虫疫苗和免疫学诊断方法的研制提供候选抗原。

The Cp23 gene of Cryptosporidium parvum was cloned into prokaryotic expression vector pET-28 a（＋）by PCR,and the recombinant expression vector pET28 a-Cp23 was constructed.The recombinant plasmid was transformed into E.coli BL21（DE3）and induced by IPTG.The expressed product was analyzed by SDS-PAGE and Western blot.The results of SDS-PAGE showed that the molecular weight of the protein was 25 000,and the protein was highly expressed as inclusion protein.The results of Western blot showed that the expressed product could be recognized by the positive serum of Cryptosporidium parvum,and had special immunogenicity.This study provides candidate antigens for the development of anti-Cryptosporidiumparvum vaccine and immunological diagnostic methods.