新型融合蛋白hEPO/TNFαM表达质粒的构建

CONSTRUCTION OF RECOMBINANT PLASMID EXPRESSING A NOVEL PROTEIN hEPO/TNFαM

目的构建hEPO/TNFαM融合毒素的表达质粒,为该融合毒素的功能研究及临床应用前景的分析评价奠定基础。方法将人促红细胞生成素(human erythropoiein,hEPO)的基因片段与人工改造的肿瘤坏死因子α(tumor necrosis factor alpha,TNFα)基因按正确的阅读框架融合,定向克隆于表达载体pET16b。结果经酶切分析及DNA序列分析,所构建的质粒均插入hEPO/TNFαM融合基因。结论利用pET16b表达载体成功构建融合毒素hEPO/TNFαM的表达质粒。

Objective To characterize the biological function and clinical perspective of hEPO/TNFαM fusion toxin,aplasmid expressing such molecule was constructed.Methods HEPO cDNA was obtained from total RNA from placenta tissue of human by RT-PCR.Artificially modified TNFα cDNA was fused precisely to the reading frame of hEPO and further cloned into the E.coli expression vector pET16b.Results The recombinant plasmid containing hEPO/TNFαM fusion gene was obtained as demonstrated by restriction analysis.Conclusion We succeed in constructing the recombinant plasmid expressing hEPO/TNFαM fusion toxin.