早期肺癌病人血液中p53的克隆及其pEGFP-N1-p53的构建  

p53 Cloning of Early Lung Cancer Patient Blood and Construction of pEGFP-N1-p53

作　　者：李振红[1] 雷雪芹[1] 徐廷生[1] 张广平[2] 韦光辉[1] 索剑飞[1] 

机构地区：[1]河南科技大学动物科技学院 [2]河南科技大学第一附属医院,河南洛阳471003

出　　处：《河南科技大学学报（自然科学版）》2011年第2期56-59,110-111,共4页Journal of Henan University of Science And Technology:Natural Science

基　　金：河南省科技攻关项目(082102130001);河南省基础与前沿技术研究计划项目(072300430010)

摘　　要：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。Human tumor suppressor gene p53 was cloned by RT-PCR from early lung cancer patient blood.Identified as wild type by sequencing,pMD18-T-p53 and the eukaryotic expression vector pEGFP-N1-p53 with enhanced green fluorescent protein(EGFP) gene were constructed by the method of digestion ligation and transformation.pEGFP-N1-p53 was digested by Hind III and BamH I,and electro-phoresis of the digested products showed two due fragments(1 188 bp and 4 695 bp).Sequencing analysis showed that the human tumor suppressor gene p53 of wild type is cloned and its eukaryotic expression vector pEGFP-N1-p53 is constructed.

关 键 词：人抗癌基因p53 克隆 真核表达载体 增强型绿色荧光蛋白 

分 类 号：Q782[生物学—分子生物学] Q819