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. 2010 Dec 21;25(6):401–408. doi: 10.1007/s12250-010-3152-y

Expression of rice gall dwarf virus outer coat protein gene (S8) in insect cells

Guo-cheng Fan 1,2, Fang-luan Gao 1,3, Tai-yun Wei 1,3, Mei-ying Huang 1,3, Li-yan Xie 1,3, Zu-jian Wu 1,3, Qi-ying Lin 1, Lian-hui Xie 1,3,✉
PMCID: PMC8227871  PMID: 21221918

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.

Key words: Rice gall dwarf virus (RGDV), Outer coat protein, Baculovirus expression system, Spodoptera frugiperda (Sf9) insect cells

Footnotes

Foundation items: This project was supported by the National Science Foundation of China (30970135), The Key Project of Genetically Modified Organisms Breeding (2009ZX08009-044B), the Natural Science Foundation of Fujian Province of China (No.2006J0065) and the Public-interest Scientific Institution Basal Research Fund of Fujian Province (2009R10029-3)

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Articles from Virologica Sinica are provided here courtesy of Wuhan Institute of Virology, Chinese Academy of Sciences

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