Biotechnology Letters

, Volume 35, Issue 3, pp 413–421

Codon optimization of the human papillomavirus type 58 L1 gene enhances the expression of soluble L1 protein in Saccharomyces cerevisiae

Original Research Paper

Abstract

The effect of codon optimization of L1 gene on the production of the L1 protein of human papillomavirus (HPV) was investigated in a yeast expression system. Saccharomyces cerevisiae was transformed with a plasmid containing either the wild type (WS)-HPV type 58 L1 (HPV58 L1) gene or a codon-optimized (MO)-HPV58 L1 gene. The proportion of soluble L1 protein expressed from MO-HPV58 L1 was significantly higher than that expressed from WS-HPV58 L1. Moreover, the amount of purified MO-HPV58 L1 protein recovered was 2.5-fold higher than the amount of WS-HPV58 L1 protein. Codon optimization of HPV58 L1 gene thus increases the proportion of soluble L1 protein and the amount of purified product that can be used as antigen to generate vaccines.

Keywords

Codon optimization Human papillomavirus L1 protein Saccharomyces cerevisiae 

Supplementary material

10529_2012_1097_MOESM1_ESM.doc (134 kb)
Supplementary material 1 (DOC 134 kb)

References

  1. Angov E (2011) Codon usage: nature’s roadmap to expression and folding of proteins. Biotechnol J 6:650–659PubMedCrossRefGoogle Scholar
  2. Bulmer M (1987) Coevolution of codon usage and transfer RNA abundance. Nature 325:728–730PubMedCrossRefGoogle Scholar
  3. Conway MJ, Meyers C (2009) Replication and assembly of human papillomaviruses. J Dent Res 88:307–317PubMedCrossRefGoogle Scholar
  4. Cook JC, Joyce JG, George HA, Schultz LD, Hurni WM, Jansen KU, Hepler RW, Ip C, Lowe RS, Keller PM, Lehman ED (1999) Purification of virus-like particles of recombinant human papillomavirus type 11 major capsid protein L1 from Saccharomyces cerevisiae. Protein Expr Purif 17:477–484PubMedCrossRefGoogle Scholar
  5. Feige MJ, Hendershot LM (2011) Disulfide bonds in ER protein folding and homeostasis. Curr Opin Cell Biol 23:167–175PubMedCrossRefGoogle Scholar
  6. Garland SM, Smith JS (2010) Human papillomavirus vaccines: current status and future prospects. Drugs 70:1079–1098PubMedCrossRefGoogle Scholar
  7. Hulsmeier AJ, Welti M, Hennet T (2011) Glycoprotein maturation and the UPR. Methods Enzymol 491:163–182PubMedCrossRefGoogle Scholar
  8. Ikemura T (1985) Codon usage and tRNA content in unicellular and multicellular organisms. Mol Biol Evol 2:13–34PubMedGoogle Scholar
  9. Kim HJ, Kim SY, Lim SJ, Kim JY, Lee SJ, Kim H-J (2010) One-step chromatographic purification of human papillomavirus type 16 L1 protein from Saccharomyces cerevisiae. Protein Expr Purif 70:68–74PubMedCrossRefGoogle Scholar
  10. Kim HJ, Kwag HL, Jin Y, Kim H-J (2011a) The composition of the carbon source and the time of cell harvest are critical determinants of the final yield of human papillomavirus type 16 L1 protein produced in Saccharomyces cerevisiae. Protein Expr Purif 80:52–60PubMedCrossRefGoogle Scholar
  11. Kim SY, Kim HJ, Kim H-J (2011b) Simple and convenient chromatography-based methods for purifying the pseudovirus of human papillomavirus type 58. Protein Expr Purif 76:103–108PubMedCrossRefGoogle Scholar
  12. Kim HJ, Lim SJ, Kwag HL, Kim H-J (2012) The choice of resin-bound ligand affects the structure and immunogenicity of column-purified human papillomavirus type 16 virus-like particles. PLoS ONE 7:e35893PubMedCrossRefGoogle Scholar
  13. Kirnbauer R, Taub J, Greenstone H, Roden R, Durst M, Gissmann L, Lowy DR, Schiller JT (1993) Efficient self-assembly of human papillomavirus type 16 L1 and L1–L2 into virus-like particles. J Virol 67:6929–6936PubMedGoogle Scholar
  14. Kozak M (2005) Regulation of translation via mRNA structure in prokaryotes and eukaryotes. Gene 361:13–37PubMedCrossRefGoogle Scholar
  15. Kudla G, Murray AW, Tollervey D, Plotkin JB (2009) Coding-sequence determinants of gene expression in Escherichia coli. Science 324:255–258PubMedCrossRefGoogle Scholar
  16. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  17. Lajoie P, Moir RD, Willis IM, Snapp EL (2011) Kar2p availability defines distinct forms of endoplasmic reticulum stress in living cells. Mol Biol Cell 23:955–964CrossRefGoogle Scholar
  18. Lindl KA, Akay C, Wang Y, White MG, Jordan-Sciutto KL (2007) Expression of the endoplasmic reticulum stress response marker, BiP, in the central nervous system of HIV-positive individuals. Neuropathol Appl Neurobiol 33:658–669PubMedCrossRefGoogle Scholar
  19. Matsukura T, Sugase M (1990) Molecular cloning of a novel human papillomarvirus (type 58) from an invasive cervical carcinoma. Virology 177:833–836PubMedCrossRefGoogle Scholar
  20. Mattanovich D, Gasser B, Hohenblum H, Sauer M (2004) Stress in recombinant protein producing yeasts. J Biotechnol 113:121–135PubMedCrossRefGoogle Scholar
  21. Park MA, Kim HJ, Kim H-J (2008) Optimum conditions for production and purification of human papillomavirus type 16 L1 protein from Saccharomyces cerevisiae. Protein Expr Purif 59:175–181PubMedCrossRefGoogle Scholar
  22. Pastrana DV, Buck CB, Pang YY, Thompson CD, Castle PE, FitzGerald PC, Kruger Kjaer S, Lowy DR, Schiller JT (2004) Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18. Virology 321:205–216PubMedCrossRefGoogle Scholar
  23. Ron D, Walter P (2007) Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 8:519–529PubMedCrossRefGoogle Scholar
  24. Soni R, Carmichael JP, Murray JA (1993) Parameters affecting lithium acetate-mediated transformation of Saccharomyces cerevisiae and development of a rapid and simplified procedure. Curr Genet 24:455–459PubMedCrossRefGoogle Scholar
  25. Webb E, Cox J, Edwards S (2005) Cervical cancer-causing human papillomaviruses have an alternative initiation site for the L1 protein. Virus Genes 30:31–35PubMedCrossRefGoogle Scholar
  26. Zhou J, Sun XY, Stenzel DJ, Frazer IH (1991) Expression of vaccinia recombinant HPV 16 L1 and L2 ORF proteins in epithelial cells is sufficient for assembly of HPV virion-like particles. Virology 185:251–257PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  1. 1.College of Pharmacy, Chung-Ang UniversitySeoulSouth Korea

Personalised recommendations