Pichia Pastoris: The Search for the Perfect Length of the 5’UTR
Poster Number
19
Format
Poster Presentation
Abstract/Artist Statement
The yeast, Pichia pastoris, has the greatest protein expression system in the world. Pichia’s success can be attributed to the Alcohol Oxidase I Promoter (AOXI) which is fused to it’s 5’ Untranslated region (UTR). The AOXI 5’UTR is used for expression of most proteins and is extremely long, consisting of 115 bases. Proteins are synthesized when DNA is transcribed into messenger RNA (mRNA) and mRNA is then translated into different proteins. There are three specific parts of mRNA: a 5’UTR, coding sequence, and a 3’UTR. The question is: can changing the number of bases in the 5’UTR increase or decrease the amount of protein expression from the RNA in P. pastoris? The goal of this project is to make deletion and insertion mutants of the AOXI 5’UTR to investigate changes in protein expression. Laboratory techniques used were mutagenesis reactions, Double Oligo Reannealing Technique (DORT) reactions, and a Lac Z reporter. To our surprise protein expression varied unexpectedly with the different mutants.
Location
Pacific Geosciences Center
Start Date
5-5-2007 1:00 PM
End Date
5-5-2007 3:00 PM
Pichia Pastoris: The Search for the Perfect Length of the 5’UTR
Pacific Geosciences Center
The yeast, Pichia pastoris, has the greatest protein expression system in the world. Pichia’s success can be attributed to the Alcohol Oxidase I Promoter (AOXI) which is fused to it’s 5’ Untranslated region (UTR). The AOXI 5’UTR is used for expression of most proteins and is extremely long, consisting of 115 bases. Proteins are synthesized when DNA is transcribed into messenger RNA (mRNA) and mRNA is then translated into different proteins. There are three specific parts of mRNA: a 5’UTR, coding sequence, and a 3’UTR. The question is: can changing the number of bases in the 5’UTR increase or decrease the amount of protein expression from the RNA in P. pastoris? The goal of this project is to make deletion and insertion mutants of the AOXI 5’UTR to investigate changes in protein expression. Laboratory techniques used were mutagenesis reactions, Double Oligo Reannealing Technique (DORT) reactions, and a Lac Z reporter. To our surprise protein expression varied unexpectedly with the different mutants.