Title

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

Lead Author Major

Biological Sciences

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

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Apr 21st, 6:00 PM Apr 21st, 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.