猪繁殖与呼吸综合征病毒非结构蛋白NSP12基因的克隆和表达

In this study, 462bp NSP12 gene of porcine resproductive and respiratory syndrome virus was cloned using RT-PCR. The NSP12 was cloned into pGEM-T vector. It was tested by sequencing and subclone into an expression vector pGEX-KG resulting in the recombinant expression plasmid pKG-NSP12. After transformed into E．coli BL21 (DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-NSP12 fusion protein was expressed about 44kDa molecular mass. Gel scanning analysis revealed 47.5% expression quantity of the fusion expression in all tropina. The result of Western blot demonstrated that the recombinant fusion protein GST-NSP12 can be recognized by anti-PRRSV monoclonal antibody. The cloning and expression of NSP12 gene of PRRSV made a foundation for the study of its function and application.