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目的 在真核细胞表达具有生物活性的可溶性肿瘤坏死因子 (TNF)受体蛋白 ,用于中和TNF的毒性。方法 将TNFR(P5 5 )的胞外区基因和人IgGFc段基因通过 18个碱基的凝血酶切点连接 ,克隆入真核表达载体pcDNA3 1(+ ) ,在COS7和BHK细胞分别得到目的蛋白的表达。结果 在真核表达系统表达了TNFR(P5 5 )胞外区与人IgGFc段的融合蛋白 ,免疫学实验及L92 9细胞体外实验均表明该蛋白具有TNFR(P5 5 )特异的抗原性、与TNF结合的活性以及抑制TNF的活性。结论 真核表达的融合蛋白产物具有可溶性TNF受体 (P5 5 )的生物学活性。
Objective To express biologically active soluble tumor necrosis factor (TNF) receptor protein in eukaryotic cells to neutralize the toxicity of TNF. Methods The extracellular domain of TNFR (P5 5) and human IgGFc gene were ligated by 18 base thrombin and cloned into the eukaryotic expression vector pcDNA3 1 (+). The target protein was obtained from COS7 and BHK cells expression. Results The fusion protein of extracellular domain of TNFR (P5 5) and human IgGFc was expressed in eukaryotic expression system. The results of immunological experiments and in vitro experiments of L92 9 cells showed that TNFR (P5 5) Binding activity as well as inhibiting TNF activity. Conclusion The eukaryotic expressed fusion protein product has the biological activity of soluble TNF receptor (P5 5).