论文部分内容阅读
首先用RT-PCR方法从早期肺癌病人的外周血中克隆出人抗癌基因p53,经测序确认为野生型后,通过酶切、连接、转化构建出扩增质粒pMD18-T-p53,然后构建出含增强型绿色荧光蛋白(EGFP)基因的真核表达质粒pEGFP-N1-p53。pEGFP-N1-P53经限制性内切酶Hind III和BamH I酶切,酶切产物电泳结果显示的两个片段(1 188 bp和4 695 bp)和实验预期相符,测序结果表明:获得野生型人抗癌基因p53并构建了真核表达载体pEGFP-N1-p53。
First, the human anti-oncogene p53 was cloned from the peripheral blood of patients with early-stage lung cancer by RT-PCR and confirmed to be wild-type by sequencing. The amplified plasmid pMD18-T-p53 was constructed by restriction enzyme digestion, ligation and transformation Eukaryotic expression plasmid pEGFP-N1-p53 containing enhanced green fluorescent protein (EGFP) gene was obtained. pEGFP-N1-P53 was digested with restriction endonucleases Hind III and BamH I, and the two fragments (1 188 bp and 4 695 bp) showed by the electrophoresis results of digestion products were consistent with the experimental ones. The sequencing results showed that the wild type Human anti-oncogene p53 and construct eukaryotic expression vector pEGFP-N1-p53.