Title

Optimization of Expression of Basic Fibroblast Growth Factor in Pichia pastoris for Oral Wound Healing

Poster Number

13C

Lead Author Major

Biological Sciences

Lead Author Status

Senior

Second Author Major

Psychology

Second Author Status

5th year Senior

Third Author Major

Biological Sciences

Third Author Status

Senior

Format

Poster Presentation

Faculty Mentor Name

Geoff Lin-Cereghino

Faculty Mentor Email

glincere@pacific.edu

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

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Apr 27th, 10:00 AM Apr 27th, 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.