TK自杀基因的构建及鉴定

目的利用分子生物学技术,构建胸苷激酶基因（TK）自杀基因并测定其结构,为该自杀基因的进一步研究提供良好的实验基础。方法以人类单纯疱疹病毒Ⅰ（HSV1）基因组DNA为模板,利用聚合酶链反应（PCR）法,扩增出约为1 100 bp TK基因,定向克隆到载体PEGM-T,构建克隆载体。两端分别引入限制性内切酶NotⅠ、XholⅠ酶切位点,经PCR及酶切鉴定初步证实为重组体后,测序认证。结果经PCR、酶切鉴定和测序鉴定完成TK基因的构建,同源性高达98.70%,完全具备表达该目的基因的要求。结论成功扩增出TK基因,为继续研究打下基础。

Construction of TK suicide gene

Objective To construct the suicide gene herpes simplex virus thymidine kinase gene(HSV-TK) and to sequence its structure for identification.Methods The TK gene fragment was amplified by PCR from the genome DNA of herpes simplex virus.The PCR product was cloned to the vector PEGM-T.The interested TK gene fragment in the recombinant plasmid obtained was confirmed by PCR,enzyme digestion and sequencing.Results The TK gene was successfully obtained.The product was 98.70% original.Conclusion The construction of TK gene may help start the research into the suicide gene strategy for the control of cancers.