Optimization of Expression of Basic Fibroblast Growth Factor in Pichia pastoris for Oral Wound Healing
Poster Number
13C
Format
Poster Presentation
Faculty Mentor Name
Geoff Lin-Cereghino
Faculty Mentor Department
Biological Sciences
Abstract/Artist Statement
One of the biggest downfalls of oral surgery is the long healing process, usually accompanied by a lot of pain. Our research aims to introduce a method of recovery that involves injecting a microbe into the oral cavity at the site of injury. We are working with Pichia pastoris in order to have it produce and secrete a recombinant human basic fibroblast growth factor (bFGF) that stimulates cellular growth. The purpose of our research is to see if yeast Pichia Pastoris can secrete recombinant proteins under mammalian tissue environment and to figure out which components in the medium: Fetal Bovine Serum (FBS) and/or Penicillin Streptomycin (P/S) promote greater protein expression in Pichia. The second part of our research focuses on the effects of mutations in the Matα gene, which is on the secretion of bFGF in Pichia pastoris.
Secretion of GFP in Pichia pastoris under yeast culture medium (BMDY) and human culture medium, Dulbecco’s Modified Eagle Media (DMEM), along with the components tested such as FBS and P/S, were compared through a Full Western. Site directed mutagenesis was used in order to create mutant strains of Pichia pastoris. The amount of bFGF secretion for each mutant strain was quantified through a Spot Western blot. The results indicate that Pichia pastoris can secrete proteins under medium mimicking mammalian tissue environment. Fetal Bovine Serum was found to be the component that increased protein secretion the greatest. The deletion of amino acids Δ 35-43 in the Matα secreted greater amounts of bFGF as compared to the wild type, super-secretor strain of Pichia pastoris. Further research still needs to be conducted to be able to put this research into practical use.
Location
DeRosa University Center Ballroom
Start Date
27-4-2018 10:00 AM
End Date
27-4-2018 12:00 PM
Optimization of Expression of Basic Fibroblast Growth Factor in Pichia pastoris for Oral Wound Healing
DeRosa University Center Ballroom
One of the biggest downfalls of oral surgery is the long healing process, usually accompanied by a lot of pain. Our research aims to introduce a method of recovery that involves injecting a microbe into the oral cavity at the site of injury. We are working with Pichia pastoris in order to have it produce and secrete a recombinant human basic fibroblast growth factor (bFGF) that stimulates cellular growth. The purpose of our research is to see if yeast Pichia Pastoris can secrete recombinant proteins under mammalian tissue environment and to figure out which components in the medium: Fetal Bovine Serum (FBS) and/or Penicillin Streptomycin (P/S) promote greater protein expression in Pichia. The second part of our research focuses on the effects of mutations in the Matα gene, which is on the secretion of bFGF in Pichia pastoris.
Secretion of GFP in Pichia pastoris under yeast culture medium (BMDY) and human culture medium, Dulbecco’s Modified Eagle Media (DMEM), along with the components tested such as FBS and P/S, were compared through a Full Western. Site directed mutagenesis was used in order to create mutant strains of Pichia pastoris. The amount of bFGF secretion for each mutant strain was quantified through a Spot Western blot. The results indicate that Pichia pastoris can secrete proteins under medium mimicking mammalian tissue environment. Fetal Bovine Serum was found to be the component that increased protein secretion the greatest. The deletion of amino acids Δ 35-43 in the Matα secreted greater amounts of bFGF as compared to the wild type, super-secretor strain of Pichia pastoris. Further research still needs to be conducted to be able to put this research into practical use.