【摘要】 目的利用重组腺相关病毒载体系统,包装含HPV16型反义E7基因重组腺相关病毒rAAV-HPV16E7AS,并进行纯化及鉴定。方法利用磷酸钙沉淀法,将腺相关病毒载体系统共转染入病毒包装细胞293细胞中,分别合成rAAV-HPV16E7AS和rAAV-lacZ,用斑点杂交法测定重组病毒的滴度,并用重组病毒感染293细胞测定其转染效率。结果所构建的rAAV-HPV16E7AS和rAAV-lacZ滴度均为1.0×1010个病毒分子/ml,平均转染效率为75%。结论已成功构建rAAV-HPV16E7AS,为临床预防和治疗子宫颈癌提供新的手段。更多还原

【Abstract】 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.更多还原