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GCRG123.方法:采用PCR技术从pGEM-T质粒上扩增出含完整ORF的GCRG123cDNA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:工程菌经IPTG诱导后,高效表达出相对分子质量约18700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23.6%.结论:在大肠杆菌中成功表达了GCRG123重组融合蛋白.
AIM： To express gastric cancer related gene GCRG123 by using thioredoxin fusion expression system.
METHODS： GCRG123 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-GCRG123 fusion protein was expressed in E.coli.
RESULTS： SDS-PAGE analysis showed the thioredoxin-GCRG123 fusion protein with relative molecule mass of 18 700 was highly expressed. The thin layer gel scanning analysis showed that the yield of GCRG123 fusion protein was 23.6% of the total bacterial protein.
CONCLUSION： The GCRG123 recombinant fusion protein is successfully expressed in E.coli.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第35期3984-3987,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~~
