Cloning of VP1 Gene of Chicken Infectious Anemia Virus and Its Expression in E.coli

We designed a pair of oligonucleotide primers based on the genomic sequence of CAV Cux-1 strain,and amplified VP1 gene by PCR.The amplification fragment was cloned into pGM-T easy vector.The result of nucleotide sequence analysis demonstrated that the cloned genomic DNA was identical with that deduced from Cux-1 strain of CAV.Then the cloned genomic DNA was subcloned into prokaryotic expression vector PET-32a(+),and the recombinant protein was expressed in E.coli cell.SDS-PAGE and Western Blot analysis showed that a protein with a molecular weight of 70.4 kDa was successfully expressed and CAV-infected chicken serum reacted with the recombinant protein.