胃癌相关基因GCRG224的克隆及其在大肠杆菌中的表达  

Cloning and expression of gastric cancer related gene GCRG224 in E.coli

作　　者：伍银桥[1] 王刚石[1] 王孟薇[1] 吴本俨[1] 尤纬缔[1] 王卫华[1] 

机构地区：[1]中国人民解放军总医院南楼消化科,北京市100853

出　　处：《世界华人消化杂志》2004年第4期763-766,共4页World Chinese Journal of Digestology

基　　金：军队"十五"重点科研基金资助项目;No.01Z035~~

摘　　要：目的:利用硫氧还蛋白融合表达系统表达胃癌相关基因GCRG224. 方法:采用PCR技术从pGEM-T质粒上扩增出含完整ORF 的GCRG224 cDNA序列,将其克隆至硫氧还蛋白融合表达载体pET102/D-TOPO中,转化大肠杆菌BL21,经IPTG 诱导表达重组融合蛋白,SDS-PAGE分析表达产物. 结果:高效表达出相对分子量约16800的重组融合蛋白.薄层凝胶扫描显示,其表达量占菌体总蛋白质的22.3%.AIM: To express gastric cancer related gene GCRG224 by using thioredoxin fusion expression system. METHODS: GCRG224 cDNA with complete open reading frame was amplified by PCR from plasmid pGEM-T, and then cloned into thioredoxin fusion expression vector pET102/D-TOPO. The recombinant plasmid was further transformed into E.coli BL21 strain. After induction with IPTG, thioredoxin-GCRG224 fusion protein was expressed in E.coli. RESULTS: SDS-PAGE analysis showed the thioredoxin-GCRG224 fusion protein with a relative molecule mass of 16 800 was highly expressed. The thin layer gel scanning analysis showed that the yield of GCRG224 fusion protein was 22.3% of the total bacterial protein. CONCLUSION: The GCRG224 recombinant fusion protein is successfully expressed in E.coli.

关 键 词：胃癌 癌基因 GCRG224 基因表达 大肠杆菌 肿瘤 

分 类 号：R735.2[医药卫生—肿瘤]