Cloning and prokaryotic expression of the E1 gene of bovine viral diarrhea virus

According to the sequence data of BVDV VEDEVAC strain published by GeneBank,one set of primer was designed and used to amplify E1 gene by the method of RT-PCR.A specific 585 bp DNA product was amplified,which was cloned into pMD18-T vector,and the positive recombinant clone was identified by restriction enzyme digestion.The recombinant plasmid was sequenced and compared with E1 gene of VEDEVAC strain.Compared with Blast on line,the E1 gene of VEDEVAC-like strain exhibits the highest homology with VEDEVAC strain,they shares 100% nucleotide sequence identity and 100% amino acid identity.It suggests that the E1 gene was phylogeneticly closer to BVDV VEDEVAC strain after phylogenetic analysis.The E1 gene was subcloned into pGEX-6p-1 expression vector,and the recombinant expression plasmid were transformed into engineering bacteria of E.coli BL21,and successfully expressed after induced.