Secretion of the Respiratory Syncytial Virus Fusion Protein From Insect Cells Using the Baculovirus Expression System

  • Boon-Huan Tan
  • Gaie Brown
  • Richard J. Sugrue
Part of the Methods in Molecular Biology book series (MIMB, volume 379)


Sequences derived from the respiratory syncytial virus (RSV) fusion (F) protein were expressed in insect cells as recombinant glutathione-S-transferase (GST)-tagged proteins. The sequence covering the F2 subunit (GST-F2), and a truncated form of the F protein in which the transmembrane domain was removed (GST-F2/F1, were cloned into the baculovirus pAcSecG2T secretory vector. These virus sequences also had the endogenous virus signal sequence removed and replaced with a signal sequence derived from the baculovirus gp67 glycoprotein, which was present in pAcSecG2T. The recombinant RSV glycoproteins were successfully detected in expressing cells by immunofluorescence assay and in the tissue culture medium by western blot analysis. The secreted recombinant GST-F2/F1 protein was further analysed using glycosidases. Our results showed that the GST-F2/F1 protein were sensitive to peptide: N-glycosidase F (PNGase F) treatment, but not to Endoglycosidase H (EndoH) treatment. This indicates that the secreted recombinant proteins were modified by the addition of mature N-linked glycan chains.

Key Words

Respiratory syncytial virus Baculovirus GST-F2 protein GST-F2/F1 protein 


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Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Boon-Huan Tan
    • 1
  • Gaie Brown
    • 2
  • Richard J. Sugrue
    • 2
  1. 1.Virology Group Defence Medical and Environmental Research InstituteDSO National LaboratoriesSingapore
  2. 2.MRC Virology UnitInstitute of VirologyGlasgowUK

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