TNF受体(P55)和IgGFc的融合蛋白在哺乳动物细胞中的表达  

Cloning and expression of human TNFR(P55)-IgG Fc fusion protein in eukaryotic cells

作　　者：安乃莉[1] 徐春晓[1] 姚立红[1] 陈爱君[1] 曾革非[1] 张立国[1] 张智清[1] 

机构地区：[1]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室,北京100052

出　　处：《中华实验和临床病毒学杂志》2001年第2期116-119,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家 8 6 3高科技生物领域基金资助项目 (10 2 0 8 0 1 0 3)

摘　　要：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 soluble biologically active TNF receptor protein in eukaryotic cells,which can inhibit cytotoxic acitivity of TNF.Methods PCR amplified the extra cellular region of TNF receptor P55 and IgG Fc gene.Then the two were linked through an oligomer encoding a thrombin sensitive peptide linker and cloned into eukaryotic expression vector pcDNA3 1(+).The eukaryotic expression plasmid pcDNA3.1?TI was then transfected to the mammalian cell line COS7 and BHK.Using the G418 system,BHK cell clones were selected and can continuously secrete biological protein in large amount.Results The expressed protein was fused with IgG Fc and secreted into the cell culture supernatant.It has good antigenicity and binding ability to TNF.It can also in hibit the cytotoxic activity of TNF on L929 cell.Conclusion The TNFR IgG Fc fusion protein expressed in eukaryotic cells has biological activities of human TNF receptor P55.

关 键 词：肿瘤坏死因子 融合蛋白质 真核表达系统 

分 类 号：R346[医药卫生—基础医学]