Construction of Chicken Infectious Anemia Virus Vector Based on VP1 and VP2 Genes

Based on the sequences of infectious anemia gene published in the Genbank,two sets of specific primers were designed and synthesized.The genes of VP1 and VP2 were amplified by PCR from the DNA extracted from the infected chicken liver tissue,and then the VP1 and VP2 genes were cloned into the pMD18-T Simple vector.Sequencing analysis showed that the sequences of VP1 and VP2 were 99.8% homology with the published sequence.The mutated mVP2 gene was obtained after VP2 gene was cloned into the pMD18-T Simple vector,site-directed mutated Vp3 initiation codon was conducted.An expression recombinant plasmid which was named pIRES-VP1-mVP2 were constructed by inserted mVP2 and VP1 into expression vector JM109.The plasmid pIRES-VP1-mVP2 was successfully obtained after extraction and enzyme digestion,which provide basis for further research of developing chichen infectous anemia vaccine.