cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells
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...
Main Author: | |
---|---|
Language: | en |
Published: |
University of Waterloo
2006
|
Subjects: | |
Online Access: | http://hdl.handle.net/10012/1234 |
id |
ndltd-LACETR-oai-collectionscanada.gc.ca-OWTU.10012-1234 |
---|---|
record_format |
oai_dc |
spelling |
ndltd-LACETR-oai-collectionscanada.gc.ca-OWTU.10012-12342014-06-18T03:51:12Z cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells Murray, Heather Biology protein localization green fluorescent protein GFP stem cells cDNA-GFP fusion library 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. 2006-08-22T14:36:03Z 2006-08-22T14:36:03Z 2005 2005 Thesis or Dissertation http://hdl.handle.net/10012/1234 en Copyright: 2005, Murray, Heather. All rights reserved. University of Waterloo |
collection |
NDLTD |
language |
en |
sources |
NDLTD |
topic |
Biology protein localization green fluorescent protein GFP stem cells cDNA-GFP fusion library |
spellingShingle |
Biology protein localization green fluorescent protein GFP stem cells cDNA-GFP fusion library Murray, Heather cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells |
description |
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. |
author |
Murray, Heather |
author_facet |
Murray, Heather |
author_sort |
Murray, Heather |
title |
cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells |
title_short |
cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells |
title_full |
cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells |
title_fullStr |
cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells |
title_full_unstemmed |
cDNA?GFP Fusion Libraries for Analyses of Protein Localization in Mouse Stem Cells |
title_sort |
cdna?gfp fusion libraries for analyses of protein localization in mouse stem cells |
publisher |
University of Waterloo |
publishDate |
2006 |
url |
http://hdl.handle.net/10012/1234 |
work_keys_str_mv |
AT murrayheather cdnagfpfusionlibrariesforanalysesofproteinlocalizationinmousestemcells |
_version_ |
1716669973023162368 |