Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Expression of Human Thrombopoietin in Insect Cells by Polyhedrin-gp64 Dual Promoter-Based Baculovirus Vector System

  • Published : 1999.06.01
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Abstract

A new baculovirus transfer vector (pPGP404) was constructed to increase the expression level of human thrombopoietin (hTPO) in insect cells. In pPGP404, hTPO was cloned next to the AcNPV polyhedrin-gp64 dual promoter and the leader sequence of hTPO was substituted with that of gp64. A recombinant baculovirus, AcPGP404, was constructed by using pPGP404 as a transfer vector. hTPO was expressed in AcPGP404-infected TN5 cells and it was observed that the expression levels of hTPO in TN5 cells increased three-fold ($6.0 {\mu}g/ml^{-1}$) compared to the level expressed under the control of the polyhedrin single promoter. These results indicate that the polyhedrin-gp64 dual promoter system would be useful for expression in large quantities of recombinant proteins in insect cells.

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References

  1. Virology v.169 Glycosylation is not required for the fusion activity of the G protein of vesicular stomatitis virus in insect cells Bailey, M. J.;D. A. McLeod;C. Y. Kang;D. H. L. Bishop
  2. J. Virol. v.65 Baculovirus gp64 gene expression: Analysis of sequences modulating early transcription and transactivation by IEI Blissard, G. W.;G. F. C. Rohrmann
  3. Nature v.369 Stimulation of megakaryocytopoiesis and thrombopoiesis by the c-mpl ligand de Sauvage, F.J.;P,E, Hass;S,D. Spencer;B.E. Malloy;A.L. Gurney;S.A. Spencer;W.C. Darbonne;W.J. Henzel;S.C. Wong;W.J. Kuang;K.J. Oles;B.Hutgren;L.A. Solberg;D.V. Goeddel;D.L. Eaton
  4. Virology v.205 Biosynthesis and processing of the Autographa californica nuclear polyhedrosis virus gp64 protein Jarvis, D. L.;A. Garcia, Jr.
  5. Mol. Cell. Biol. v.9 Glycosylation and secretion of human tissue plasminogen activator in recombinant baculovirus-infected cells Jarvis, D. L.;M. D. Summers
  6. J. Microbiol. v.34 Analysis of fusogenic activity of Autographa california nuclear polyhedrosis virus (AcNPV) gp64 glycoprotein Kim, H. J.;J. M. Yang
  7. J. Biochem. Mol. Biol. v.31 Secretory production of biologically active human thrombopoietin by baculovirus expression system Koh, Y.W.;S. W. Lim;S. K. Park;M. H. Park;D. S. Na;J. M. Yang
  8. Nature v.369 Cloning and expression of murine thrombopoietin cDNA and stimulation of platelet production in vivo Lok, S.;K. Kaushansky;R. D. Holly;J. L. Kuijper;C. E. Lofton-Day;P. J. Oort;F. J. Grant;M. D. Heipel;S. K. Burkhead;J. M. Kramer;L. A. Bell;C. A. Sprecher;H. Blumberg;R. Jonson;D. Prunkard;A. F. T.Ching;S. L. Mathewes;M. C. Bailey;J. W. Forstrom;M. M. Buddle;S. G. Osborn;S. J. Evans;P. O. Sheppard;S. R. Prensnell;P. J. O'Hara;F. S. Hagen;G. Roth;D. C. Foster
  9. Gen. Anal. Tech. Appl. v.7 Temporal expression of HIV-I envelope proteins in baculovirus-infected insect cells: Implications for glycosylation and CD4 binding Murphy, C. I.;M. Lennick;S. M. Lehar;G. A. Beltz;E. Young
  10. Protein Expr. Purif. v.4 Enhanced expression, secretion, and large-scale purification of recombinant HIV-1 gp 120 in insect cell using the baculovirus egt and p67 signal peptides Murphy, C. I.;J. R. McIntirel;D. R. Davis;H. Hodgdon;J. R. Seals;E. Young
  11. Baculovirus Expression Protocols Richardson, C. D.
  12. Gene v.131 Differential secretion and glycosylation of recombinant human chorionic gonadotropin (beta hCG) synthesized using different promoters in the baculovirus expression vector system Sridhar, P.;S. E. Hasnain
  13. Gene v.98 Enhanced secretion from insect cells of a foreign protein fused to the honeybee melittin signal peptide Tessier. D. C.;D. Y. Thomas ;H. E. Khouri;F. Laliberte;T. Vernet
  14. Gene v.91 Differential gene expression mediated by late, very late and hybrid baculovirus promoters Thiem, S. M.;L. K. Miller
  15. J. Virol. v.63 Identification and sequence analysis of a gene encoding gp67, an abundant envelope glycoprotein of the baculovirus Autographa californica nuclear polyhedrosis virus Whiteford, M.;S. Stewart;J. Kuzio;P. Faulkner