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目的克隆表达细粒棘球绦虫亲环蛋白(EgCyP)基因,并对其进行生物信息学分析。方法从细粒棘球绦虫cD NA中扩增目的基因,克隆入表达载体pET28a,转化大肠埃希菌(E.coli)BL21(DE3),经异丙基βD硫代半乳糖苷诱导表达后,进行SDS PAGE和免疫印迹试验鉴定,并对其进行生物信息学分析。结果重组质粒pET28a EgCyP构建成功。SDS PAGE和免疫印迹试验结果显示,重组蛋白在E.coliBL21(DE3)中获得高效表达,重组蛋白EgCyP分子量约为22 kDa,可被细粒棘球绦虫感染犬血清识别。生物信息学分析显示该蛋白具有7个潜在的抗原表位。结论成功克隆出细粒棘球绦虫EgCyP基因并在E.coliBL21(DE3)中表达,为进一步研究其免疫原性奠定了基础。
Objective To clone and express the gene encoding Echinococcus granulosus (CYP9C), and analyze its bioinformatics. Methods The target gene was amplified from cD NA of Echinococcus granulosus and cloned into the expression vector pET28a. The recombinant plasmid was transformed into E. coli BL21 (DE3). After induced by isopropyl βD thiogalactoside, SDS PAGE and Western blot test identification, and its bioinformatics analysis. Results The recombinant plasmid pET28a EgCyP was successfully constructed. SDS PAGE and Western blotting results showed that the recombinant protein was highly expressed in E.coli BL21 (DE3). The recombinant protein EgCyP has a molecular weight of about 22 kDa and can be recognized by the canine serum of Echinococcus granulosus. Bioinformatics analysis revealed that the protein has seven potential epitopes. Conclusion The EgCyP gene of Echinococcus granulosus was successfully cloned and expressed in E.coli BL21 (DE3), which laid the foundation for further study of its immunogenicity.