AI Article Synopsis
- Pichia pastoris is a genetically modified yeast used to produce over a thousand different proteins valuable for various fields like industry and medicine.
- Researchers studied the alcohol oxidase 1 (AOX1) gene's 5' untranslated region (UTR) to see how its length affects protein expression.
- They found that changing the 5'UTR length influences how efficiently proteins are made, revealing that it has both positive and negative factors that complicate efforts to create an optimized version for better expression.
Article 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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3464313 | PMC |
| http://dx.doi.org/10.1016/j.gene.2012.01.006 | DOI Listing |
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