胃癌相关基因GCRG213的克隆及其在大肠杆菌中的表达  

Cloning and expression of gastric cancer related gene GCRG213 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]解放军总医院南楼消化科,北京100853

出　　处：《军医进修学院学报》2004年第4期258-260,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：军队"十五"重点科研基金项目资助 ( 0 1Z0 3 5 )

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GCRG2 13。方法 :采用PCR技术从pGEM T质粒上扩增出含完整ORF的GCRG2 13cDNA序列 ,将其克隆至硫氧还蛋白融合表达载体 pET10 2 /D TOPO中 ,转化大肠杆菌BL2 1,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2 9.4ku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 2 8.7%。结论 :在大肠杆菌中成功表达了Thioredoxin GCRG2 13融合蛋白 ,为后续功能研究、蛋白制备及其抗体研制奠定了基础。Objective:To express gastric cancer related gene GCRG213 using thioredoxin fusion expression system. Methods:GCRG213 cDNA with complete open reading frame was amplified by PCR from plasmid pGEM-T, and then was cloned into thioredoxin fusion expression vector pET102/D-TOPO. The recombinant plasmid was further transformed into E.coli BL21 strain. After induction with IPTG, GCRG213 fusion protein was expressed in E.coli. Results:SDS-PAGE analysis showed the thioredoxin-GCRG213 fusion protein with relative molecule mass of 29.4ku was highly expressed. The thin layer gel scanning analysis showed that the yield of GCRG213 fusion protein was 28.7% of the total bacterial protein. Conclusion:The thioredoxin-GCRG213 fusion protein was successfully expressed in E.coli. This might provide an experimental basis for the function research, protein and antibody preparation hereafter.

关 键 词：胃癌 GCRG213基因 基因克隆 大肠杆菌 基因表达 

分 类 号：R735.2[医药卫生—肿瘤]