Rabies virus glycoprotein expression in Drosophila S2 cells. I. Functional recombinant protein in stable co-transfected cell line. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biotechnology Journal Année : 2007

Rabies virus glycoprotein expression in Drosophila S2 cells. I. Functional recombinant protein in stable co-transfected cell line.

Adriana y Yokomizo
  • Fonction : Auteur
Soraia a C Jorge
  • Fonction : Auteur
Renato M Astray
  • Fonction : Auteur
Irene Fernandes
  • Fonction : Auteur
Orlando G Ribeiro
  • Fonction : Auteur
Denise S P Q Horton
  • Fonction : Auteur
Aldo Tonso
  • Fonction : Auteur
Noel Tordo

Résumé

Recombinant rabies virus glycoprotein (rRVGP) was expressed in Drosophila melanogaster Schneider 2 (S2) cells. The cDNA encoding the entire RVGP gene was cloned in an expression plasmid under the control of the constitutive actin promoter (Ac), which was co-transfected into S2 cells together with a hygromycin selection plasmid. Selected S2 cell populations (S2AcRVGP) had a decreased ability to grow and consume substrates, when compared to the non-transfected cells (S2). They were shown, by PCR, to express the RVGP gene and mRNA and, by immunoblotting, to synthesize the rRVGP in its expected molecular mass of 65 kDa. ELISA kinetic studies showed the rRVGP expression in cell lysates and supernatants attaining concentrations of 300 microg/L. By flow cytometry analysis, about 30% of the cells in the co-transfected populations were shown to express the rRVGP. Cell populations selected by limiting dilution expressed higher rRVGP yields. Mice immunized with rRVGP were shown to synthesize antibodies against rabies virus and be protected against experimental infection with rabies virus. The data presented here show that S2 cells can be suitable hosts for the rRVGP expression, allowing its synthesis in a high degree of physical and biological integrity.

Domaines

Virologie

Dates et versions

hal-00167792 , version 1 (23-08-2007)

Identifiants

Citer

Adriana y Yokomizo, Soraia a C Jorge, Renato M Astray, Irene Fernandes, Orlando G Ribeiro, et al.. Rabies virus glycoprotein expression in Drosophila S2 cells. I. Functional recombinant protein in stable co-transfected cell line.. Biotechnology Journal, 2007, 2 (1), pp.102-9. ⟨10.1002/biot.200600211⟩. ⟨hal-00167792⟩

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