摘要
目的构建胃癌新生血管导向性肽GX1与新型人肿瘤坏死因子(GX1-rmhTNF)基因的原核表达载体,表达GX1-rmhTNF蛋白。方法利用基因工程方法将胃癌血管特异性结合肽融合在新型人肿瘤坏死因子α的氨基末端,构建GX1-rmhTNF原核表达载体,在大肠杆菌进行表达。对表达产物进行SDS-PAGE电泳分析和Westernblot检测分析。结果成功构建了GX1-rmhTNF重组融合表达质粒,表达的蛋白经SDS-PAGE电泳分析,在相对分子质量(Mr)约18000处出现了1条新生的蛋白条带,该表达蛋白具有与TNFα单克隆抗体特异性的结合能力。结论成功构建了GX1-rmhT-NF基因的原核表达载体并在原核细胞中表达了该产物,为下一步GX1-rmhTNF的纯化奠定了基础。
AIM： To construct prokaryotic vector GX1-rmhTNF containing neovascular-targeting peptide GX1 and human TNFα and produce the GX1-rmhTNF protein. METHODS： We have previously obtained a specific phage peptide CGNSNPKSC （ GX1 ） binding to vasculature of human gastric cancer. The GX1-rmhTNFα vector was constructed by merging sequence of neovascular-targeting peptide GX1 with N-terminal of new recombined human TNFα （rmhTNFα） using gene engineering methods. The expression of the fusion protein GXI-rmhTNFα in E. coli was induced by temperature. The expression of GX1-rmhTNFα was detected by SDS-PAGE and Western blot. RESULTS： A novel protein with expected molecular mass about 18 000 was found after SDS-PAGE and gel staining. The expressed product showed a good binding ability to anti-TNFα monoclonal antibody. CONCLUSION： The prekaryotic vector GX1-rmhTNF was constructed and the expression of GX1-rmhTNF protein was successfully induced, which was helpful for further purification of GX1-rmhTNF protein.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2006年第3期360-362,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(No.30130260
30572148)
军队科技攻关课题(No.01Z087)
