Date of Award

10-2011

Degree Type

Thesis

Degree Name

Master of Science in Biomedical Sciences

First Advisor

Quinn Lu, PhD

Second Advisor

Richard Kriebel, PhD

Third Advisor

Susan T Hingley, PhD

Abstract

Baculovirus has been a common tool for protein production since the mid 1980s. The virus has an exclusive tropism only capable of infecting the larval stages of insects. Originally, baculovirus recombinant gene expression used Spodoptera frugiperda (Sf9) insect cells as the host cells for protein production. However, in 1995, it was discovered that baculoviruses could be modified to transduce mammalian cells with the insertion of a mammalian promoter and gene of interest. Due to its ease of generation and low biology safety hazards, this technology, known as BacMam (Baculovirus Mammalian) is popular in the research field for protein production and for cell based and membrane binding drug discovery assays. BacMam viruses have shown high transduction efficiencies in many widely used mammalian cell lines such as CHO-K1, HEK293F, and U-2 OS cells. However, some cancer cell lines do not transduce well. Because disease relevant cell lines are preferred in cell based assays, we want to increase the transduction efficiencies in these lines. Pseudotyping, a way of modifying the wild type envelope protein to enhance target specificity, could be a method of increasing efficiencies. We have generated four pseudotyping viruses using the Vesicular Stomatitis Virus glycoprotein (VSV-G).

COinS