马尔堡病毒糖蛋白GP1亚基的真核表达与纯化  

Eukaryotic Expression and Purification of Marburg Virus Envelope GP1 Protein

作　　者：张冠英 迟象阳 范鹏飞 房婷 吴诗坡 陈旖 王美荣 陈郑珊 于长明 陈薇 

机构地区：[1]军事科学院军事医学研究院生物工程研究所,北京100071

出　　处：《生物技术通讯》2018年第1期17-20,共4页Letters in Biotechnology

基　　金：国家科技重大专项(2014ZX09J14301)

摘　　要：目的:用真核表达系统可溶性表达重组马尔堡病毒糖蛋白GP1亚基并纯化。方法:从糖蛋白(GP)全长基因上扩增GP1基因序列,克隆至真核表达载体pcDNA3.4,在哺乳动物表达系统中进行表达,镍柱亲和层析纯化目的蛋白,ELISA检测重组蛋白与特异抗体的免疫反应性。结果:GP1在哺乳动物表达系统中获得可溶性表达,纯化得到的目的蛋白与特异抗体具有良好的结合活性,推测其构象与天然马尔堡病毒GP1具有相似性。结论:真核表达了马尔堡病毒糖蛋白GP1亚基,且目的蛋白具有良好的抗原性,可用于GP特异抗体筛选、疫苗效果评价等研究。Objective: To construct an expressing vector of Marburg virus(MARV) envelope GP1 protein, and express recombinant GP1 protein in mammalian expression system. Methods: The GP1 sequence was amplified from full-length gene of MARV glycoprotein(GP) and cloned into pc DNA3.4 vector. After sequencing, the recombinant pc DNA3.4-GP1 was transfected into Expi293 cells. The protein of interest was purified through Ni affinity chromatography and identified by Western blot and ELISA assays. Results: We obtained recombinant GP1 protein from the supernatant of transfected Expi293 cells. Purified GP1 protein can bind anti-MARV GP specific monoclonal antibodies. Conclusion: GP1 protein has been expressed in mammalian expression system, and can be used to screen specific antibodies and evaluate vaccine efficacy.

关 键 词：马尔堡病毒 糖蛋白 GP1亚基 Expi293细胞 

分 类 号：Q78[生物学—分子生物学]