Clone And Prokaryotic Expression Of VP4 Gene Of Bovine Rotavirus

Bovine rotaviruses are nonenveloped,segmented,doublestranded RNA viruses classified in the family Reoviridae. Bovine rotaviruses are the major cause of diarrhea in the piglets.Bovine rotavirus has been widely distributed in the world and cost pig producers serious loss.In order to construct the prokaryotic expression vector pET32a -VP4, total RNA were extracted form MA104 cell infected with bovine rotavirus G6 strain CHLY isolated by our laboratory.The open reading frame of VP4 (2331bp) was amplified by RT-PCR and cloned into pEASY-T3 vector using T-A cloning technique VP4 gene was subcloned into the expression vector pET32a and transformed into E. coli BL21 (DE3). SDS-PAGE showed that the bacterium induced by IPTG successfully expressed, approximately 108 KD exogenous protein was observed on the SDS- PAGE. The results indicate that the fusion protein is of antigenic expected to lay foundation for further studies of the subunit vaccine and DNA vaccine and to prevention of bovine rotavirus.