Display options
Cite
Martínez-Solís M, Gómez-Sebastián S, Escribano JM, et al. A novel baculovirus-derived promoter with high activity in the baculovirus expression system. PeerJ. 2016;4:e2183doi: 10.7717/peerj.2183.
Martínez-Solís, M., Gómez-Sebastián, S., Escribano, J. M., Jakubowska, A. K., & Herrero, S. (2016). A novel baculovirus-derived promoter with high activity in the baculovirus expression system. PeerJ, 4e2183. https://doi.org/10.7717/peerj.2183
Martínez-Solís, María, et al. "A novel baculovirus-derived promoter with high activity in the baculovirus expression system." PeerJ vol. 4 (2016): e2183. doi: https://doi.org/10.7717/peerj.2183
Martínez-Solís M, Gómez-Sebastián S, Escribano JM, Jakubowska AK, Herrero S. A novel baculovirus-derived promoter with high activity in the baculovirus expression system. PeerJ. 2016 Jun 28;4:e2183. doi: 10.7717/peerj.2183. eCollection 2016. PMID: 27375973; PMCID: PMC4928464.
Copy Download .nbib Format: NLM
Share it on

PeerJ. 2016 Jun 28;4:e2183. doi: 10.7717/peerj.2183. eCollection 2016.

A novel baculovirus-derived promoter with high activity in the baculovirus expression system.

PeerJ

María Martínez-Solís, Silvia Gómez-Sebastián, José M Escribano, Agata K Jakubowska, Salvador Herrero

Affiliations

  1. Department of Genetics, Universitat de València, Burjassot, Spain; Estructura de Recerca Interdisciplinar en Biotecnologia i Biomedicina (ERI BIOTECMED), Universitat de València, Burjassot, Valencia, Spain.
  2. Alternative Gene Expression S.L. (ALGENEX) , Madrid , Spain.
  3. Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) , Madrid , Spain.
  4. Department of Genetics, Universitat de València , Burjassot , Spain.

PMID: 27375973 PMCID: PMC4928464 DOI: 10.7717/peerj.2183

Abstract

The baculovirus expression vector system (BEVS) has been widely used to produce a large number of recombinant proteins, and is becoming one of the most powerful, robust, and cost-effective systems for the production of eukaryotic proteins. Nevertheless, as in any other protein expression system, it is important to improve the production capabilities of this vector. The orf46 viral gene was identified among the most highly abundant sequences in the transcriptome of Spodoptera exigua larvae infected with its native baculovirus, the S. exigua multiple nucleopolyhedrovirus (SeMNPV). Different sequences upstream of the orf46 gene were cloned, and their promoter activities were tested by the expression of the GFP reporter gene using the Autographa californica nucleopolyhedrovirus (AcMNPV) vector system in different insect cell lines (Sf21, Se301, and Hi5) and in larvae from S. exigua and Trichoplusia ni. The strongest promoter activity was defined by a 120 nt sequence upstream of the ATG start codon for the orf46 gene. On average, GFP expression under this new promoter was more than two fold higher than the expression obtained with the standard polyhedrin (polh) promoter. Additionally, the orf46 promoter was also tested in combination with the polh promoter, revealing an additive effect over the polh promoter activity. In conclusion, this new characterized promoter represents an excellent alternative to the most commonly used baculovirus promoters for the efficient expression of recombinant proteins using the BEVS.

Keywords: Insect virus; Nucleopolyhedrovirus; Protein expression

References

  1. J Biotechnol. 2013 Jun 10;165(3-4):201-8 - PubMed
  2. Cell Biol Toxicol. 2010 Feb;26(1):57-68 - PubMed
  3. J Virol. 2010 Jul;84(14):7233-42 - PubMed
  4. Arch Virol. 1996;141(7):1247-58 - PubMed
  5. J Virol. 1992 Dec;66(12):7397-405 - PubMed
  6. J Virol. 2013 Jun;87(11):6391-405 - PubMed
  7. J Biol Chem. 2002 Feb 15;277(7):5256-64 - PubMed
  8. J Gen Virol. 1991 Feb;72 ( Pt 2):275-83 - PubMed
  9. J Biotechnol. 2014 Mar 10;173:41-6 - PubMed
  10. J Gen Virol. 1994 May;75 ( Pt 5):1115-23 - PubMed
  11. Virology. 1993 Jan;192(1):273-81 - PubMed
  12. Insect Biochem Mol Biol. 2012 Aug;42(8):557-70 - PubMed
  13. Methods Enzymol. 2009;463:191-222 - PubMed
  14. Biochem Biophys Res Commun. 2004 Oct 15;323(2):599-614 - PubMed
  15. J Gen Virol. 2015 Jan;96(Pt 1):6-23 - PubMed
  16. PLoS One. 2014 May 13;9(5):e96562 - PubMed
  17. Nucleic Acids Res. 1988 May 11;16(9):3635-53 - PubMed
  18. FEBS Lett. 2002 Dec 4;532(1-2):143-6 - PubMed
  19. Virology. 1988 Nov;167(1):56-71 - PubMed
  20. Mol Cell Biol. 1983 Dec;3(12):2156-65 - PubMed
  21. Biochemistry. 2004 Jun 29;43(25):8143-51 - PubMed
  22. J Mol Biol. 1989 Dec 20;210(4):721-36 - PubMed
  23. Virol Sin. 2012 Apr;27(2):71-82 - PubMed
  24. Biotechnol Lett. 2010 Nov;32(11):1637-47 - PubMed
  25. Methods. 2011 Sep;55(1):52-7 - PubMed
  26. Biotechnol Bioeng. 2010 Dec 15;107(6):909-16 - PubMed
  27. Gene. 1990 Jul 2;91(1):87-94 - PubMed
  28. J Gen Virol. 1991 Oct;72 ( Pt 10):2551-6 - PubMed
  29. J Biotechnol. 2013 May 10;165(1):11-7 - PubMed
  30. J Gen Virol. 1987 May;68 ( Pt 5):1233-50 - PubMed
  31. Arch Virol. 1990;111(1-2):103-14 - PubMed
  32. Mol Biotechnol. 2010 Sep;46(1):80-9 - PubMed

Publication Types