Construction and Expression of the Recombinant Adenovirus Expressing the NS3 Antigen of Hepatitis C Virus

To develop an attenuated recombinant adenoviral vaccine against Hepatitis C virus infection , we construct the recombinant adenoviral vector expressing non-structural protein 3(NS3) antigen of HCV(RAd-NS3). The HCV NS3 gene enconding for 329-935aa was amplified from the plasmid pRC/NS3 by PCR and cloned into the adenoviral shuttle plasmid pAdTrack-CMV. A two-step transformation protocol was employed for the construction of recombinant adenoviral plasmid pAdEasy-HCV NS3. The identified recombinant DNA was transfected into 293 cells to package adenovirus. From cell lysis , the presence of recombinant adenovirus was continued by PCR, and RT-PCR demonstrated the transcription of NS3 in HepG2 cells infected with RAd-NS3, and Western blot revealed that NS3 protein can express in infected HepG2 cells. These results indicated that tumor cells infected with recombinant adenovirus vector RAd-NS3 can express NS3 antigen ,which will be potentially used as a candidate vaccine for the prophylaxis and