关键词: 小鼠细小病毒, VP2 蛋白, 杆状病毒表达 

Abstract: Objective Recombinant MVM VP2 protein was synthesized by baculovirus expression system and the immunogenicity of the recombinant VP2 protein was analyzed. Method The optimized MVM VP2 gene sequence was cloned into the eukaryotic expression vector pFastBac1, then the recombinant vector pFastBac1-MVM VP2 was transformed into DH10Bac susceptible E. coli to form recombinant plasmid. The recombinant plasmid were transfected into insect cells Sf9. Recombinant baculovirus rBacMVM VP2 was harvested after significant cellular lesions were observed. Sf9 cells were inoculated with recombinant virus rBac-MVM VP2 for 48 and 72 h. Immunogenicity of the recombinant protein was tested by IFA and Western blot. Result Recombinant baculovirus rBac-MVM VP2 could successfully express recombinant MVM VP2 protein after infecting Sf9 cells. Western blot and IFA analysis showed that the recombinant protein had good immunogenicity and high expression at 72 h of virus infection. After protein purification, a high purity recombinant protein VP2 was obtained. Conclusion In this study, MVM VP2 protein was successfully expressed by insect cell-baculovirus system and the recombinant VP2 protein has good immunogenicity, which lays a foundation for the study of VP2-related functions and the establishment of clinical diagnosis method.

Key words: minute virus of mice, VP2 protein, baculovirus expression