UWSpace is currently experiencing technical difficulties resulting from its recent migration to a new version of its software. These technical issues are not affecting the submission and browse features of the site. UWaterloo community members may continue submitting items to UWSpace. We apologize for the inconvenience, and are actively working to resolve these technical issues.
 

cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells

Loading...
Thumbnail Image

Date

2005

Authors

Murray, Heather

Journal Title

Journal ISSN

Volume Title

Publisher

University of Waterloo

Abstract

Stem cells have great potential value for treating a number of diseases and conditions, including diabetes, Parkinson's, and spinal cord injuries. Applying stem cells for therapeutic purposes will require an in-depth understanding of their biology, not only of the genes they express, but also the functions of the proteins encoded by the genes. The goal of the project presented in this thesis was to develop a method for high-throughput analyses of protein localization in mouse stem cells. Localization information can provide insight into the functions and biological roles of proteins. <br /><br /> One means of studying protein localization involves creating proteins with a green fluorescent protein (GFP) reporter gene and analyzing their localization using fluorescence microscopy. The research outlined in this thesis focused on developing a system to create a large number of GFP-tagged proteins by constructing a cDNA?GFP fusion library. This involved exploring methods for optimizing cDNA synthesis, designing a retroviral vector (pBES23) for the expression of cDNA?GFP fusions in mouse stem cells, and constructing a cDNA?GFP fusion library in this vector using R1 mouse embryonic stem cell mRNA. The library constructed was not successfully delivered to target cells for GFP-tagged protein expression; it was therefore not possible to characterize protein localization in mouse stem cells. Suggestions are given as to how the methods used in this thesis might be optimized further.

Description

Keywords

Biology, protein localization, green fluorescent protein, GFP, stem cells, cDNA-GFP fusion library

LC Keywords

Citation

Collections