稳定表达BmCPV病毒VP4蛋白受体细胞系的建立

Bombyx mori Cytoplasmic Polyhedrosis virus, which is called BmCPV for short and belongs to Cypovirus of Reoviridae, is double-stranded RNA virus. As the one of Bombyx mori's main toxic pathogen, it usually does great harm to sericultural production. So it will be great of significance to prevent utilize BmCPV to conduct biological centrol through go into the mechanism of interacton between BmCPV and Bombyx mori. Acquire the fusion gene called yfpn-vp4 through overlap PCR, .After linking vp4 gene with pMD19-T Simple Vector and getting plasmid TS-yfpn-vp4. After that, TS-yfpn-vp4 and pIZTV5-His are digested by restriction endonuclease, ligation and transform into DH5αin order to get recombinant plasmid pIZTV5-His-yfpn-vp4. Then transfect recombinant plasmid pIZTV5-His-yfpn-vp4 into BmN cells, Zeocin is used for screening transfected BmN cells. It turned out that green fluorescence can be observed 72 hours after transfection. At the same time, the results of SDS-PAGE and Western Blot both show that fusion protein YFPN-VP4 can be stably expressed in vitro. The above research results not only laid a solid foundation for the interaction between BmCPV and host cells, but also provide a new method for utilizing silkworm cells to express exogenous genes.