Analysis of the 5’Untranslated Region (5’UTR) of the Alcohol Oxidase 1 Gene as a Regulator of Translation in Pichia pastoris
Poster Number
8
Format
Poster Presentation
Faculty Mentor Name
Joan Lin-Cereghino
Faculty Mentor Department
Biological Sciences
Additional Faculty Mentor Name
Geoff Lin-Cereghino
Abstract/Artist Statement
Introduction: Pichia pastoris is a yeast commonly used for foreign protein expression. The coding sequences of foreign proteins are inserted after the AOXI promoter. The 5’ untranslated region (UTR) is part of the mRNA before the coding sequence, which affects the rate of translation (protein production) by ribosomes.Objective: We are trying to figure out the correlation between 5’ UTR structure and degree of protein expression.Methods: Oligonucleotide primers were used in mutagenesis to make deletions in the 5’ UTR on a plasmid that contains the beta-galactosidase gene (encoding a reporter protein) as the coding sequence. We used beta-galactosidase assays to measure protein expression in the yeast.Results and Conclusions: All deletions caused decreased beta-galactosidase expression in the yeast, suggesting that they all enhance translation. No negative-acting sequences have ever been found.
Location
DeRosa University Center, Ballroom
Start Date
21-4-2011 6:00 PM
End Date
21-4-2011 8:00 PM
Analysis of the 5’Untranslated Region (5’UTR) of the Alcohol Oxidase 1 Gene as a Regulator of Translation in Pichia pastoris
DeRosa University Center, Ballroom
Introduction: Pichia pastoris is a yeast commonly used for foreign protein expression. The coding sequences of foreign proteins are inserted after the AOXI promoter. The 5’ untranslated region (UTR) is part of the mRNA before the coding sequence, which affects the rate of translation (protein production) by ribosomes.Objective: We are trying to figure out the correlation between 5’ UTR structure and degree of protein expression.Methods: Oligonucleotide primers were used in mutagenesis to make deletions in the 5’ UTR on a plasmid that contains the beta-galactosidase gene (encoding a reporter protein) as the coding sequence. We used beta-galactosidase assays to measure protein expression in the yeast.Results and Conclusions: All deletions caused decreased beta-galactosidase expression in the yeast, suggesting that they all enhance translation. No negative-acting sequences have ever been found.