Title

Characterization of the Bgs13 protein in Pichia pastoris super-secretion

Poster Number

03B

Lead Author Major

Biological Sciences, Pre-Dentistry

Lead Author Status

Senior

Second Author Major

Biological Sciences

Second Author Status

Senior

Third Author Major

Biological Sciences, Pre-Dentistry

Third Author Status

Sophomore

Format

Poster Presentation

Faculty Mentor Name

Geoffrey Lin-Cereghino

Faculty Mentor Email

glincere@pacific.edu

Faculty Mentor Department

Biological Sciences

Additional Faculty Mentor Name

Joan Lin-Cereghino

Additional Faculty Mentor Email

jlincere@u.pacific.edu

Additional Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

The yeast Pichia pastoris is known to be efficient at expressing and secreting recombinant proteins, including human insulin and a hepatitis B vaccine protein. Pichia pastoris has been known to secrete certain proteins well while struggling to secrete others. Secreted proteins are easier to purify and therefore are more useful than non-secreted proteins. Our lab has created a strain that is a super-secretor, with a mutated Bgs13 (Beta-galactosidase supersecretion) gene. In order to determine why bgs13 is a super-secretor, cell wall assays were performed. Congo red and Calcoflour white assays were used to determine if super-secretion is a result of defective cell walls. In addition, the Bgs13 protein is also known to have protein kinase C activity. We are testing if super-secretion is a result of elevated or decreased protein kinase C activity. Lastly, localization of wild type Bgs13 protein and mutant bgs13 protein were compared by fusing each protein to EGFP, and their location was determined with fluorescence microscopy analysis. By characterizing the mutant and wild type Bgs13 proteins using these three methods, the results will help us create strains with optimized secretion of many different recombinant proteins.

Location

DeRosa University Center, Ballroom

Start Date

29-4-2017 10:00 AM

End Date

29-4-2017 12:00 PM

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Apr 29th, 10:00 AM Apr 29th, 12:00 PM

Characterization of the Bgs13 protein in Pichia pastoris super-secretion

DeRosa University Center, Ballroom

The yeast Pichia pastoris is known to be efficient at expressing and secreting recombinant proteins, including human insulin and a hepatitis B vaccine protein. Pichia pastoris has been known to secrete certain proteins well while struggling to secrete others. Secreted proteins are easier to purify and therefore are more useful than non-secreted proteins. Our lab has created a strain that is a super-secretor, with a mutated Bgs13 (Beta-galactosidase supersecretion) gene. In order to determine why bgs13 is a super-secretor, cell wall assays were performed. Congo red and Calcoflour white assays were used to determine if super-secretion is a result of defective cell walls. In addition, the Bgs13 protein is also known to have protein kinase C activity. We are testing if super-secretion is a result of elevated or decreased protein kinase C activity. Lastly, localization of wild type Bgs13 protein and mutant bgs13 protein were compared by fusing each protein to EGFP, and their location was determined with fluorescence microscopy analysis. By characterizing the mutant and wild type Bgs13 proteins using these three methods, the results will help us create strains with optimized secretion of many different recombinant proteins.