[Expression of hepatitis E virus ORF3 gene fragment in baculovirus system and its immunological character]
1Department of Molecular Biology, Institute of Medical Biology, CAMS and PUMC, Kunming 650118, China.
Objective:
To express hepatitis E virus (HEV) ORF3 protein by baculovirus system and provide basis for immunological character research.
Methods:
Hepatitis E Virus ORF3 gene fragment was obtained by RT-PCR, ligated with vector pThioHisA for sequencing and then inserted into transfected vector pVL1393 to construct recombinant plasmid. Mediated by Lipofectin Reagent, the recombinant vector and baculovirus linearized DNA (BaculoGold) co-transfected insect cell Sf9 to make recombinant baculovirus. Expressed ORF3 was analyzed for its immunological character by Western blotting, and immunized Kunming Mice.
Results:
Recombinant ORF3 protein could be recognized by the known positive serum and promoted organism to produce HEV-specific antibody.
Conclusions:
Recombinant baculovirus can express effectively HEV ORF3, which has HEV specific immunogenic character.
