Characterization of the Bgs13 protein in Pichia pastoris super-secretion
Poster Number
03B
Format
Poster Presentation
Faculty Mentor Name
Geoffrey Lin-Cereghino
Faculty Mentor Department
Biological Sciences
Additional Faculty Mentor Name
Joan Lin-Cereghino
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
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.