title: [Characterization and development of recombinant vaccinia viruses expressing different segments of spike protein derived from human coronavirus NL-63].
abstract: The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.
authors.affiliation: Institute of Medical Virology, Wenzhu Medical College, Wenzhu 325000, China.
authors.forename: Guo-Xia
authors.initials: GX
authors.lastname: Zhao
authors.forename: Wei-Min
authors.initials: WM
authors.lastname: Zhou
authors.forename: Rou-Jian
authors.initials: RJ
authors.lastname: Lu
authors.forename: Hui-Juan
authors.initials: HJ
authors.lastname: Wang
authors.forename: Min
authors.initials: M
authors.lastname: Zhao
authors.forename: Ting-Yin
authors.initials: TY
authors.lastname: Zhang
authors.forename: Yao
authors.initials: Y
authors.lastname: Deng
authors.forename: Ji-Ming
authors.initials: JM
authors.lastname: Gao
authors.forename: Wen-Jie
authors.initials: WJ
authors.lastname: Tan
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D058957:Coronavirus NL63, Human / Q000737:chemistry*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis*; D064370:Spike Glycoprotein, Coronavirus; D014616:Vaccinia virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins