逆转录病毒载体介导双靶区乙型肝炎病毒反义RNA模型的构建

To probe the ways by which the antisense gene of HBV can be transcribed in eukaryotic cell to inhibit HBV replication and expression, the recombinant retrovirus vector carrying dual target antisense RNA of hepatitis B virus was constructed. Antisenses of HBV subtype ayw X region and P region were constructed by artificial synthesis and were named the X and P fragments.The X and P fragments were inserted into the retrovirus pLXSN vector cloning site in the sense or antisense orientation respectively with recombinogenic technique.Then recombinant retrovirus vector carrying sing target antisense RNA was constructed successfully.Construction of recombinant retrovirus vector carrying dual target antisense RNA of HBV is in the foundation of construction of recombinant retrovirus vector carrying sing target in similar ways.The recombinant retroviurs vector carrying dual target antisense RNA of hepatitis B virus had been constructed successfully and it had been authenticated by restriction enzyme digestion and electrophoretic analysis fragments of.PCR amplification and DNA sequence code were checked with automatic sequenator.The recombinant retrovirus vector carrying dual target antisense RNA of hepatitis B virus will be a good foundation for studying the inhibition of variable HBV in eukaryotic cell and for the antivirus treatment of polygenic disease.