- Collection:
- Atlanta University and Clark Atlanta University Theses and Dissertations
- Title:
- Expression of the mouse dihydrofolate reductase gene, 2000
- Creator:
- Alvelo Ceron, Demaris
- Date of Original:
- 2000-05-01
- Subject:
- Degrees, Academic
Dissertations, Academic - Location:
- United States, Georgia, Fulton County, Atlanta, 33.749, -84.38798
- Medium:
- theses
- Type:
- Text
- Format:
- application/pdf
- Description:
- Yeast Artificial Chromosomes (YACs) containing the mouse DHFR gene were isolated, mapped and transferred into DHFR deficient Chinese Hamster Ovary (DHFR'DEFICIENT CHO) cells. The steady state levels of mouse DHFR mRNA species were analyzed during resting and growth stimulation, in an unamplified cell line. Growth arrest was obtained by serum deprivation, and growth stimulation by subsequent serum restoration. Analysis of the cell cycle was performed by Flow Cytometry. We have examined the expression and regulation of endogenous DHFR mRNA in 3T3 cells and observed a 2 fold increase in DHFR mRNA levels in cells predominantly in S phase (growing for 22H after serum stimulation) vs. arrested cells. Also, we have observed a 2 fold increase in DHFR mRNA levels in CHO cells containing the mouse DHFR gene, cultured under the same conditions. These results indicated that the transferred YAC gene is expressed and regulated in a growth dependent manner and at levels nearly identical to the endogenous 3T3 DHFR gene. We concluded that transfer of a YAC containing the mouse DHFR gene into an established DHFR'DEFICIENT CHO cell line, results in statistical comparable expression of the mouse DHFR gene. We concluded that this is an excellent system to study the expression of the mouse DHFR gene, with complete gene sequences.
- External Identifiers:
- Metadata URL:
- http://hdl.handle.net/20.500.12322/cau.td:2000_alvelo_ceron_demaris
- Rights Holder:
- Clark Atlanta University
- Holding Institution:
- Atlanta University Center Robert W. Woodruff Library
- Rights:
-
