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对腺病毒－单纯疱疹病毒胸苷激酶基因（ＡＤＶ－ＨＳＶ－ＴＫ）治疗载体进行构建与鉴定。利用脂质体介导的共转染，将质粒ＰＪＭ１７与ＦＰＡ／ＴＫ－ＲＳＶ共转染入２９３细胞系，利用挑取单噬斑，获取单克隆的重组腺病毒，并在２９３细胞系中进行扩增、纯化。经ＰＣＲ、ＤＮＡ测序，酶切鉴定，用空斑实验（ｐｌａｑｕｅａｓｓａｙ）测定，分析病毒的活性。结果：重组病毒经ＰＣＲ测序证实已携带ＴＫ片段，酶切鉴定出现特异性的酶切图谱，重组病毒具高滴度的感染活性。结论：ＡＤＶ－ＴＫ基因的构建为ＨＳＶ－ＴＫ基因的临床应用提供了更为有效的手段。
The adenovirus-herpes simplex virus thymidine kinase gene (ADV-HSV-TK) vector was constructed and identified. Plasmid PJM17 and FPA / TK-RSV were co-transfected into 293 cell line by liposome-mediated cotransfection. Monoclonal recombinant adenovirus was obtained by single plaque picking and was performed in 293 cell line Amplification, purification. After PCR, DNA sequencing, restriction enzyme digestion, plaque assay (plaqueassay) assay, analysis of the virus activity. Results: The recombinant virus was confirmed by PCR sequencing to carry the TK fragment. The specific digestion pattern was identified by restriction enzyme digestion and the recombinant virus was highly titered. Conclusion: The construction of ADV-TK gene provides a more effective method for the clinical application of HSV-TK gene.