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构建含CMV启动子和HSV-TK基因的复制缺陷型重组腺病毒载体(AdCMVTK),用PCR及Southern杂交筛选鉴定阳性空斑,证实AdCMVTK含HSV-TK基因。纯化的Ad-CMVTK滴度达10~(12)pfu/ml。GCV对经Ad-CMVTK感染(M.O.I.=100)的HeLa等3种肿瘤细胞的IC50<4μmol/L,同时观察到明显的旁杀伤效应。AdCMVTK/GCV系统的建立为肿瘤基因治疗的研究提供了新的手段。
A replication-defective recombinant adenoviral vector (AdCMVTK) containing the CMV promoter and the HSV-TK gene was constructed, and positive plaques were identified by PCR and Southern hybridization to confirm that AdCMVTK contained the HSV-TK gene. The purified Ad-CMVTK titer reached 10~12 pfu/ml. The IC50 of GCV against HeLa and other tumor cells infected with Ad-CMVTK (M.O.I.=100) was less than 4 μmol/L. At the same time, a clear side-effect was observed. The establishment of the AdCMVTK/GCV system provides a new tool for the study of tumor gene therapy.