Cloning of VP1 and 3D Gene of Duck Hepatitis Virus 1 (DHV1) and Its Expression in Escherichia coli

Abstract:

In this study, two special primer pair containing EcoR V and Xho I according to complete genome of duck hepatitis virus 1 (DHV1) were designed to amplify VP1 and 3D genes from cDNA of DHV1. The target genes VP1 and 3D were subcloned into PET32a vector digested by EcoR V and Xho I respectively. Then the recombinant plasmids were transfected into Escherichia coli BL21(DE3) for VP1and 3D expression. The bacteria containing PET32a-VP1 and PET32a-3D were collected and examined by SDS-PAGE and western-blotting. Result showed that the VP1 and 3Dprotein were expressed in E. coli and the amount of expression was higher. Molecular weight of the protein was 48 kD, 68 kD. The protein can be recognized by DHV1antibody. This study showed that the protein VP1 and 3D have antigenicity.

Key words:Duck hepatitis virus 1, VP1gene, 3Dgene, Cloning and expression