title: A new strategy for full-length Ebola virus glycoprotein expression in E.coli.
abstract: Ebola virus (EBOV) causes severe hemorrhagic fever in humans and non-human primates with high rates of fatality. Glycoprotein (GP) is the only envelope protein of EBOV, which may play a critical role in virus attachment and entry as well as stimulating host protective immune responses. However, the lack of expression of full-length GP in Escherichia coli hinders the further study of its function in viral pathogenesis. In this study, the vp40 gene was fused to the full-length gp gene and cloned into a prokaryotic expression vector. We showed that the VP40-GP and GP-VP40 fusion proteins could be expressed in E.coli at 16 °C. In addition, it was shown that the position of vp40 in the fusion proteins affected the yields of the fusion proteins, with a higher level of production of the fusion protein when vp40 was upstream of gp compared to when it was downstream. The results provide a strategy for the expression of a large quantity of EBOV full-length GP, which is of importance for further analyzing the relationship between the structure and function of GP and developing an antibody for the treatment of EBOV infection.
authors.affiliation: Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.|University of Chinese Academy of Sciences, Beijing, 100049, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Zai
authors.affiliation: Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.|University of Chinese Academy of Sciences, Beijing, 100049, China.
authors.forename: Yinhua
authors.initials: Y
authors.lastname: Yi
authors.affiliation: Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
authors.forename: Han
authors.initials: H
authors.lastname: Xia
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China. yzm@wh.iov.cn.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Yuan
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D029043:Ebolavirus / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D061987:Sf9 Cells; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C080981:VP40 protein, virus; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; C113189:envelope glycoprotein, Ebola virus
keywords: E.coli; Ebola virus (EBOV); glycoprotein (GP); prokaryotic expression