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

  • 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GCRG213。方法: 采用PCR技术从pGEM T质粒上扩增出含完整ORF的GCRG213cDNA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。

     

    Abstract: 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.

     

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