title: Expression of rice gall dwarf virus outer coat protein gene (S8) in insect cells.
abstract: To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity, its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system. The S8 gene was subcloned into the pFastBac™1 vector, to produce the recombinant baculovirus transfer vector pFB-S8. After transformation, pFB-S8 was introduced into the competent cells (E. coli DH10Bac) containing a shuttle vector, Bacmid, generating the recombinant bacmid rbpFB-S8. After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection, Sf9 cells were collected at different times and analyzed by SDS-PAGE, Western blotting and immunofluorescence microscopy. The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells. Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.
authors.affiliation: Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Guo-cheng
authors.initials: GC
authors.lastname: Fan
authors.forename: Fang-luan
authors.initials: FL
authors.lastname: Gao
authors.forename: Tai-yun
authors.initials: TY
authors.lastname: Wei
authors.forename: Mei-ying
authors.initials: MY
authors.lastname: Huang
authors.forename: Li-yan
authors.initials: LY
authors.lastname: Xie
authors.forename: Zu-jian
authors.initials: ZJ
authors.lastname: Wu
authors.forename: Qi-ying
authors.initials: QY
authors.lastname: Lin
authors.forename: Lian-hui
authors.initials: LH
authors.lastname: Xie
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D008856:Microscopy, Fluorescence; D012275:Oryza; D012087:Reoviridae / Q000235:genetics*; D018411:Spodoptera
chemical_list: D036022:Capsid Proteins