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目的:评价以轮状病毒(RV)重组VP6蛋白为载体插入Ⅱ型脊髓灰质炎病毒(PV2)VP1蛋白上的1个抗原表位构建而成的嵌合蛋白的体外免疫学性质。方法:采用分子克隆和基因重组技术将PV2抗原表位插入到RV载体蛋白上,在大肠杆菌中表达并用SDS-PAGE确认表达产物,再通过动物免疫、Western blot、免疫荧光和病毒血清抗体中和试验分析嵌合蛋白的免疫学性质。结果:成功构建了以VP6为载体的PV2抗原表位嵌合蛋白6F/PV2N1,并且在E.coli系统中高效表达,嵌合蛋白免疫的豚鼠血清抗体对RV和PV2具备较好的中和活性。结论:以RV VP6为载体构建的嵌合蛋白具有较好的免疫原性,免疫豚鼠产生血清抗体可中和RV和PV2在体外细胞上的感染;进一步为研发RV/PV2嵌合疫苗提供了较好的基础。
AIM: To evaluate the in vitro immunological properties of a chimeric protein constructed by inserting an epitope on the VP1 protein of type 2 poliovirus (PV2) with the recombinant VP6 protein of rotavirus (RV) as a carrier. Methods: PV2 epitope was inserted into RV vector by molecular cloning and gene recombination technique, expressed in E. coli and confirmed by SDS-PAGE, and then identified by animal immunization, Western blot, immunofluorescence and virus serum antibody neutralization Experimental analysis of the immunological properties of chimeric proteins. Results: The PV6 antigen-epitope chimeric protein 6F / PV2N1 was successfully constructed and highly expressed in E. coli system. The chimeric protein-immunized guinea pig serum antibody had good neutralizing activity on RV and PV2 . CONCLUSION: The chimeric protein with RV VP6 as carrier has good immunogenicity. Immunization of guinea pig with serum antibody can neutralize the infection of RV and PV2 on cells in vitro. Good foundation