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Article
Analysis of the 5' untranslated region (5'UTR) of the alcohol oxidase 1 (AOX1) gene in recombinant protein expression in Pichia pastoris
Gene
  • Chris A. Staley, University of the Pacific
  • Amy Huang, University of the Pacific
  • Maria Nattestad, University of the Pacific
  • Kristin T. Oshiro, University of the Pacific
  • Laura E. Ray, University of the Pacific
  • Tejas Mulye, University of the Pacific
  • Zhiguo Harry Li, Novo-Nordisk Pharmaceuticals Science and Technology
  • Thu Le, University of the Pacific
  • Justin J. Stephens, University of the Pacific
  • Seth R. Gomez, University of the Pacific
  • Allison D. Moy, University of the Pacific
  • Jackson C. Nguyen, University of the Pacific
  • Andreas H. Franz, University of the Pacific
  • Joan Lin-Cereghino, University of the Pacific
  • Geoff P. Lin-Cereghino, University of the Pacific
Document Type
Article
Department
Chemistry
DOI
10.1016/j.gene.2012.01.006
Publication Date
4-1-2012
Disciplines
Abstract
Pichia pastoris is a methylotrophic yeast that has been genetically engineered to express over one thousand heterologous proteins valued for industrial, pharmaceutical and basic research purposes. In most cases, the 5′ untranslated region (UTR) of the alcohol oxidase 1 (AOX1) gene is fused to the coding sequence of the recombinant gene for protein expression in this yeast. Because the effect of the AOX1 5′UTR on protein expression is not known, site-directed mutagenesis was performed in order to decrease or increase the length of this region. Both of these types of changes were shown to affect translational efficiency, not transcript stability. While increasing the length of the 5′UTR clearly decreased expression of a β-galactosidase reporter in a proportional manner, a deletion analysis demonstrated that the AOX1 5′UTR contains a complex mixture of both positive and negative cis-acting elements, suggesting that the construction of a synthetic 5′UTR optimized for a higher level of expression may be challenging.
Citation Information
Chris A. Staley, Amy Huang, Maria Nattestad, Kristin T. Oshiro, et al.. "Analysis of the 5' untranslated region (5'UTR) of the alcohol oxidase 1 (AOX1) gene in recombinant protein expression in Pichia pastoris" Gene Vol. 496 Iss. 2 (2012) p. 118 - 127 ISSN: 0378-1119
Available at: http://works.bepress.com/andreas-franz/97/