Date of Graduation

5-2015

Document Type

Thesis

Degree Name

Bachelor of Science in Biological Engineering

Degree Level

Undergraduate

Department

Biological and Agricultural Engineering

Advisor/Mentor

Jin, Sha

Committee Member/Reader

Carrier, Danielle J.

Committee Member/Second Reader

Wochok, Jeffrey C.

Abstract

Influenza viruses are enveloped viruses with segmented RNA genome surrounded by a helical symmetry shell. Due to genetic reassortment between avian, swine, and human influenza viruses, a global pandemic may emerge, calling for new methods of mass vaccine production. Yeast is an ideal organism to express viral antigens (e.g. hemagglutinin) because of its natural adjuvant activity, making the expressed proteins more immunogenic when administered to the human body [1,2]. In addition to the purpose of vaccination, yeast is economically convenient by expressing proteins in a fast, inexpensive manner [3,4]. In this study, the goal is to express the HA1 fragment of the hemagglutinin protein from swine influenza (H1N1) virus [A/Cali/4/09] on the surface of yeast stain S. cerevisiae using a reconstructed shuttle vector, pYD5. Development of recombinant yeast cells consisted of ligating genetic sequence of HA1 into pYD5, transforming into E.coli cells, and electroporating DNA plasmids into EBY100 S. cerevisiae cells. Primary antibody for the flu strain was used in conjunction with a fluorescing secondary antibody, allowing visual analysis under a microscope. In result, surface expression of HA1 fragment was demonstrated by immunofluorescence microscopy. This study represents the first steps in the generation of yeast-based vaccines for the protection of influenza viruses.

Keywords

swine flu; vaccine; flu virus; epidemiology

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