人内皮抑素基因的质粒构建及在大肠杆菌中的表达  

HUMAN ENDOSTATIN:ITS HIGN LEVEL EXPERSSION IN E COLI

作　　者：郝志芳[1] 谈立松[2] 殷智榕[1] 曲建 沈琼 

机构地区：[1]河南医科大学第一附属医院病理科,郑州450052 [2]上海市肺科医院肺癌研究室

出　　处：《广西医科大学学报》2000年第5期811-813,共3页Journal of Guangxi Medical University

摘　　要：目的 :将人内皮抑素 (endostatin)基因插入表达载体并进行原核表达。方法 :将 endostatin基因定向插入表达载体 p QE-31Bam H I/ Kpn I位点 ,构建重组质粒 p QEN,转化 E.coli M15 ,进行原核表达。结果 :在 IPTG的诱导下 ,endostatin基因在重组转化株 E.coli M15 (p QEN)中获得高效表达 ,SDS- PAGE显示表达产物分子量为 2 2× 10 3u,表达产物主要以包涵体形式存在 ,表达量占菌体总蛋白的 34 .6 %。结论 :从组织中克隆Objective:To obtain recombinant human endostatin by a prokaryotic expression system Methods:Human endostatin gene was cloned on BamH I/Kpn I site of the expression vector pQE 31 to result in the recombinant plasmid pQEN The pQEN was transformed into E coli M15 and was expressed by this prokaryoticexpression system Results:Endostatin was efficiently expressed in recombinant strain E coli M15(pQEN) as inclusion body by IPTG induction The expression product constitutes about 34 6%of the total bacterial protein and was characterized by SDS PAGE to have molecular weights of 22×10 3 u Conclusion:The method of cloning endostatin gene from human fatal liver and express it in a prokaryotic expression system are feasible and useful.

关 键 词：人内皮抑素 大肠杆菌 原核表达 质粒构建 肿瘤 

分 类 号：R730.5[医药卫生—肿瘤] Q78[医药卫生—临床医学]