Cloning and Prokaryotic Expression of VP3 Gene of Duck Hepatitis A Virus Type 1 Subtype

The VP3 gene of the hepatitis A virus type 1subtype(DHAV-1a)in ducks was amplified by the reverse transcription-polymerase chain reaction(RT-PCR)using one pair of specific primers designed according to the published sequences of DHAV-1a.The target DNA was purified and cloned into pEASYTM-Blunt Zero Cloning Vector.The recombinant expression plasmid,pET-32a-VP3,was constructed by inserting the target gene fragment into pET-32a(+)vector and transformed into Escherichia coli BL21(DE3)competent cells.In this study,SDSPAGE and Western-blot analyses showed that the recombinant protein VP3,approximately 47 kDa in molecular mass,was expressed highly in E.coli after pET-32a-VP3 was induced with 1.0mmol·L-1 IPTG at 37℃.The expressed recombinant protein was recognized specifically by Anti-His Mouse mAb showing agood bioactivity.