作 者：
王中一;李佳明;付莹莹;郭振东;陆兵;钱军;刘林娜
关键词：
埃博拉病毒病;糖蛋白;真核表达;Protein A亲和层析
摘 要：
埃博拉病毒糖蛋白与病毒黏附、入侵宿主细胞等过程密切相关,为了更好地研究该糖蛋白与宿主细胞的相互作用,拟通过哺乳动物细胞表达系统,构建GP1亚基Fc标签融合蛋白。经鉴定,成功构建了埃博拉病毒糖蛋白GP1亚基的真核表达载体pFUSE-GP1-hIgG1-Fc2,并利用该载体及293T细胞构建了表达GP1亚基融合蛋白的哺乳动物细胞系。该融合蛋白N端带有IL2分泌信号肽,C端带有人IgG1-Fc2标签,可在293T细胞以分泌表达的方式存在于细胞培养上清中,而后利用Protein A亲和层析进行融合蛋白纯化。经Western Blots鉴定,成功制备GP1-Fc融合蛋白。这不仅为埃博拉病毒糖蛋白的研究提供了技术支撑,也为其他种类蛋白的哺乳动物细胞真核表达纯化提供了新思路。
译 名：
Eukaryotic Expression and Purification of Ebola Virus Glycoprotein GP1 Subunit Fusion Protein
作 者：
WANG Zhongyi;LI Jiaming;FU Yingying;GUO Zhendong;LU Bing;QIAN Jun;LIU Linna;Academy of Military Medical Sciences, Academy of Military Sciences;College of Animal Science and Technology,Jilin Agricultural University;
单 位：
WANG Zhongyi%LI Jiaming%FU Yingying%GUO Zhendong%LU Bing%QIAN Jun%LIU Linna%Academy of Military Medical Sciences, Academy of Military Sciences%College of Animal Science and Technology,Jilin Agricultural University
关键词：
Ebola virus;;glycoprotein;;eukaryotic expression;;Protein A affinity chromatography
摘 要：
Ebola virus glycoprotein(EBOV_GP) plays important roles in viral Adhesion invasion in host cells. In order to explore the interaction effects of EBOV-GP in host cells, GP1 subunit fusion protein was constructed by mammalian cell expression system in this study. It was identified that eukaryotic expression vector pFUSE-GP1-hIgG1-Fc2 was constructed and 293 T cell lines expressing GP1-Fc were established for producing fusion protein. The fusion protein GP1-Fc, which has IL-2 secreting signal peptides in its N-terminal and a IgG1-Fc2 tag in its C-terminal, can be expressed in 293 T cells by secreting into the cell culture supernatant. GP1-Fc fusion protein was successfully prepared by Protein A affinity chromatography for purification and identified by Western Blots. This study could not only provide a technical support for other studies on EBOV_GP, but also show a new perspective for other eukaryotic expression and purification of fusion proteins.
