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23
phenotypes. The traditional tet-on system was constructed to confirm these phenotypes
on transgenic lines when the re-tests were performed.
Materials and Methods
Drosophila strains and culture
Drosophila strains and culture conditions are as previously described (Ford, Hoe et al.
2007). Experiments were preformed at 25oC using a standard cornmeal/agar media
(Ren, Webster et al. 2007). To cause conditional gene expression using the Tet-on
system, flies were cultured on media containing doxycycline and ampicillin each at a
final concentration of 64ug/ml (“+DOX”). For the control groups (“-DOX”), flies were
cultured on media containing ampicillin only.
Generation of wild type female fecundity curve
A cohort of 40 Oregon-R virgin female flies was collected over a period of 48 hours and
designated as 1 day old. They were separated into 10 vials with 4 virgins in each vial
together with 4 young Oregon-R males and kept at 25oC. Flies in each vial were
transferred to fresh food vials every second day and the number of dead flies was
recorded, and the old vials were kept at 25oC for subsequent progeny counts. Each
month 4 young Oregon-R males were added to each vial to replenish the old males, and
Object Description
| Title | Characterization of Drosophila longevity and fecundity regulating genes |
| Author | Li, Yishi |
| Author email | yishili@usc.edu; yishili@gmail.com |
| Degree | Doctor of Philosophy |
| Document type | Dissertation |
| Degree program | Molecular & Computational Biology |
| School | College of Letters, Arts and Sciences |
| Date defended/completed | 2008-08-19 |
| Date submitted | 2008 |
| Restricted until | Unrestricted |
| Date published | 2008-10-31 |
| Advisor (committee chair) | Tower, John |
| Advisor (committee member) |
Finkel, Steven E. Aparicio, Oscar Martin Longo, Valter D Comai, Lucio |
| Abstract | The regulation of Drosophila melanogaster longevity and fecundity involves many factors. Longevity is governed by oxidative stress, stem cell loss, dietary restriction, the insulin/IGF-1 pathway, and other factors. Fecundity is also regulated by multiple tissues and factors, including the germline stem cells and stem cell niche, the fat body, yolk proteins, and sex peptides. The fecundity of wild type female Drosophila gradually declines during aging, suggesting a common pathway regulating longevity and fecundity machinery. Since both mechanisms involve multiple factors, sorting through the Gordian’s knot is a formidable task. Using a PdL mutagenesis approach, I screened for a specific phenotype in thousands of independent mutant strains to examine both regulatory networks simultaneously. Two novel genes, magu and hebe, were identified and characterized to regulate longevity and fecundity. While Drosophila lifespan was extended upon the induction of these genes, fecundity increase requires that the gene induction be in an ideal range to show the expected phenotypic change. I also performed several other projects, including studying the lifespan extension effect of dIAP2, characterization of a Drosophila gut driver strain, and intra-abdominal RNAi injection in adult Drosophila. These projects provided us insight on longevity, fecundity, anti-apoptosis, stem cell biology, RNAi and other aspects of Drosophila research. In sum, Drosophila melanogaster, as a model organism for molecular biology and genetics study, will continue to contribute to the scientific community. |
| Keyword | Drosophila; longevity; fecundity |
| Language | English |
| Part of collection | University of Southern California dissertations and theses |
| Publisher (of the original version) | University of Southern California |
| Place of publication (of the original version) | Los Angeles, California |
| Publisher (of the digital version) | University of Southern California. Libraries |
| Provenance | Electronically uploaded by the author |
| Type | texts |
| Legacy record ID | usctheses-m1735 |
| Contributing entity | University of Southern California |
| Rights | Li, Yishi |
| Repository name | Libraries, University of Southern California |
| Repository address | Los Angeles, California |
| Repository email | cisadmin@lib.usc.edu |
| Filename | etd-Li-2382 |
| Archival file | uscthesesreloadpub_Volume44/etd-Li-2382.pdf |
Description
| Title | Page 33 |
| Contributing entity | University of Southern California |
| Repository email | cisadmin@lib.usc.edu |
| Full text | 23 phenotypes. The traditional tet-on system was constructed to confirm these phenotypes on transgenic lines when the re-tests were performed. Materials and Methods Drosophila strains and culture Drosophila strains and culture conditions are as previously described (Ford, Hoe et al. 2007). Experiments were preformed at 25oC using a standard cornmeal/agar media (Ren, Webster et al. 2007). To cause conditional gene expression using the Tet-on system, flies were cultured on media containing doxycycline and ampicillin each at a final concentration of 64ug/ml (“+DOX”). For the control groups (“-DOX”), flies were cultured on media containing ampicillin only. Generation of wild type female fecundity curve A cohort of 40 Oregon-R virgin female flies was collected over a period of 48 hours and designated as 1 day old. They were separated into 10 vials with 4 virgins in each vial together with 4 young Oregon-R males and kept at 25oC. Flies in each vial were transferred to fresh food vials every second day and the number of dead flies was recorded, and the old vials were kept at 25oC for subsequent progeny counts. Each month 4 young Oregon-R males were added to each vial to replenish the old males, and |
