Package,Purification and Identification of Recombinant Adeno-associated Virus rAAV-HPV16E7AS Containing Antisense E7 Gene of Human Papillomavirus 16

Objective To package,purify and identify the recombinant adeno-associated virus(rAAV)containing the antisense E7 gene of human papillomavirus 16(HPV16)by using rAAV vector system.Methods Co-transfect virus packaging cells(293 cells)with rAAV vector system by calcium phosphate precipitation to synthesize rAAV-HPV16E7AS and rAAV-lacZ respectively.Determine the titers of recombinant viruses by dot hybridization.Transfect 293 cells with the recombinant viruses and determine the transfection efficiency.Results Both the titers of constructed rAAV-HPV16E7AS and rAAV-lacZ were 1.0×1010 molecules/ml,and their mean transfection efficiency was 75%.Conclusion rAAV-HPV16E7AS was successfully constructed,which provided a novel route to prevention and treatment of cervical cancer.