胃癌相关基因GCRG213的克隆及其在大肠杆菌中的表达

目的：利用硫氧还蛋白融合表达系统表达胃癌相关基因GCRG213。方法：采用PCR技术从pGEM-T质粒上扩增出含完整ORF的GCRG213 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29.4ku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28.7％。结论：在大肠杆菌中成功表达了Thioredoxin-GCRG213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。

Cloning and expression of gastric cancer related gene GCRG213 in E.coli

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.