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Cytotechnology

, Volume 65, Issue 5, pp 829–838 | Cite as

Kinetic studies of recombinant rabies virus glycoprotein (RVGP) cDNA transcription and mRNA translation in Drosophila melanogaster S2 cell populations

  • R. M. AstrayEmail author
  • S. A. C. Jorge
  • M. A. N. Lemos
  • A. Y. Yokomizo
  • V. L. L. Boldorini
  • A. L. P. Puglia
  • O. G. Ribeiro
  • C. A. PereiraEmail author
Original Research

Abstract

Recombinant rabies virus glycoprotein (RVGP) was expressed in cell membranes of stably transfected Drosophila S2 cells using constitutive and inducible promoters. Although with quantitative differences of RVGP expression in both systems, the cDNA transcription, as evaluated by relative RVGP mRNA levels measured by qRT-PCR, sustained the amount of RVGP producing cells and the RVGP volumetric (ΠRVGP) productivity. At the transition to the stationary cell growth phase, once the cell culture slowed down its rate of multiplication, an accumulation of RVGP mRNA and RVGP was clearly observed in both cell populations. Nevertheless, cell cultures performed under sub-optimal temperatures indicated that an envisaged increase in the RVGP production is not only dependent on cell growth rate, but essentially on optimal cell metabolic state.

Keywords

S2 cells Rabies virus glycoprotein qRT-PCR mRNA Cell culture 

Abbreviations

RVGP

Rabies virus glycoprotein

qRT-PCR

Quantitative reverse transcriptase-polimerase chain reaction

X

Cell concentration (cell/mL)

Pc

Rabies virus glycoprotein cell content (μg/107 cells)

μ

Specific cell growth rate (h−1)

ΠRVGP

Volumetric productivity (ng mL−1 h−1)

Notes

Acknowledgments

We thank Dr. Jorge Mário da Costa Ferreira Jr., Laboratório de Imunoquímica, Instituto Butantan, São Paulo, SP, Brazil, for flow cytometry analysis. This work was financially supported by grants from FAPESP (2011/08331-0), CNPq and Butantan Foundation. C. A. Pereira is recipient of a CNPq 1A senior fellowship.

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • R. M. Astray
    • 1
    Email author
  • S. A. C. Jorge
    • 1
  • M. A. N. Lemos
    • 1
  • A. Y. Yokomizo
    • 1
    • 2
  • V. L. L. Boldorini
    • 1
  • A. L. P. Puglia
    • 1
  • O. G. Ribeiro
    • 3
  • C. A. Pereira
    • 1
    Email author
  1. 1.Laboratório de Imunologia ViralInstituto ButantanSão PauloBrazil
  2. 2.Cristália Produtos Químicos e Farmacêuticos LtdaItapiraBrazil
  3. 3.Laboratório de ImunogenéticaInstituto ButantanSão PauloBrazil

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