Study Of Antiviral Effects On Hepatitis B Virus With The Recombinant Adenovirus AdMxA

PARTâ… CLONING AND SEQUENCE ANALYSIS OF HUMAN MxA GENEObjective To clone the human MxA gene from the human peripheral blood mononuclear cells (PBMCs) in order to construct the recombinant adenovirus AdMxA.Methods The peripheral blood mononuclear cells(PBMCs) were isolated by gradient fractionation,total RNA was extracted from the PBMCs.The MxA gene was amplified by RT-PCR with the primers based on the published sequence of MxA.The PCR product, double-digested with restrictive endonucleases Not I and Xba I after purified, was cloned into the adenovirus shuttle plasmid(pAdTrack-CMV) by molecular cloning technique. The recombinant plasmid was confirmed by restriction analysis and sequencing.Results The restrictive endonuclease analysis confirmed that correct recombinant plasmid pAdTrack-MxA was constructed successfully. Sequencing analysis revealed that the amino acid sequence of MxA gene had just one amino acid residue which was displaced from isoleucine to...