摘要
目的:构建hTERT启动子启动的双自杀基因载体CDTK(pc-ChCDTK)。方法:利用PCR扩增CMV增强子、hTERT启动子、yCD及TKgly4种元件,并构建过渡载体T-CMV,T-hTERT,T-yCD,T-TKgly,连至pc-DNA3.1(-)质粒载体上构建成双自杀基因载体pc-DNA3.1-CMV-hTERT-CDTK(pc-ChCDTK);对各个元件、过渡载体及最终载体进行琼脂糖凝胶电泳、酶切和测序鉴定。结果:实验所构建的最终载体的酶切产物琼脂糖凝胶电泳所得片段大小分别为6773bp和288bp,与预期片段一致;测序证实最终载体与pc-ChCDTK序列一致。结论:成功构建了含有hTERT启动子的肿瘤特异性双自杀基因载体pc-ChCDTK。
AIM： To construct the tumor cells specific vector of suicide gene of CDglyTK driven by hTERT promoter. METHODS： The CMV enhancer, hTERT promoter, yCD and TKgly gene were amplified with polymerase chain reaction （PCR） technique. The transient vectors, T-CMV, T-hTERT, T-yCD, T-TKgly, were established. These fragment were inserted into plasmid vector pcDNA3.1 and aquired plasmid vector pc-bNA3.1-CMV- hTERT - CDTK （pc-ChCDTK）. The vector was identified by enzyme cutting and sequencing, RESULTS ：We got 6 773bp and 288bp fragments through enzyme cutting. The result of sequencing was equal to the expection. CONCLUSION： The vector that contains CDTK fusion gene was constructed successfully.
出处
《国际眼科杂志》
CAS
2009年第10期1876-1880,共5页
International Eye Science
