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CHORION GENE AMPLIFICATION IN DROSOPHILA MELANOGASTER
by
Nan Chen
—————————————————————————————
A Dissertation Presented to the
FACULTY OF THE GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the Requirements for the Degree
DOCTOR OF PHILOSOPHY
(MOLECULAR BIOLOGY)
August 2010
Copyright 2010 Nan Chen
Object Description
| Title | Chorion gene amplification in Drosophila melanogaster |
| Author | Chen, Nan |
| Author email | nanchen@usc.edu; nanchen@gmail.com |
| Degree | Doctor of Philosophy |
| Document type | Dissertation |
| Degree program | Molecular Biology |
| School | College of Letters, Arts and Sciences |
| Date defended/completed | 2010-04-23 |
| Date submitted | 2010 |
| Restricted until | Unrestricted |
| Date published | 2010-06-24 |
| Advisor (committee chair) | Tower, John |
| Advisor (committee member) |
Finch, Caleb E. Turcotte, Lorraine |
| Abstract | Drosophila chorion genes are amplified in the ovarian follicle cells during oogenesis in order to satisfy the high demand for eggshell protein synthesis. Amplification occurs through repeated firing of origin(s) and is strictly regulated both temporally and spatially. The third chromosome chorion gene s18 requires upstream replicator Amplification Element on the 3rd Chromosome (ACE3) and downstream sequence-specific origin ori-beta for efficient amplification. Using constructs carrying ACE3, s18 and ori-beta, we were able to study the sequence and transcription requirement for s18 amplification. We found that experimentally reducing transcription of s18 inhibits amplification. Using conditional overexpression of Green Fluorescence Protein (GFP), we found that active transcription alone is not sufficient to initiate amplification without the presence of ACE3 and ori-beta. Point mutations in the s18 TATA box reduced s18 amplification. We therefore conclude that transcription, although not sufficient for amplification initiation, is essential for efficient s18 amplification. Point mutation of ecdysone receptor binding motifs found in the s18 promoter and in ori-beta reduced both chorion gene transcription and amplification. Reduced amplification was also observed with conditional inhibition of ecdysone receptor gene expression and upon overexpression of dominant-negative forms of the ecdysone receptor. Ecdysone receptor therefore mediates both developmental regulation of chorion gene transcription and amplification. Additionally, we identified several novel positive and negative trans-regulators of chorion gene amplification, via a gene over-expression screen. Taken together, the results demonstrate that chorion gene amplification is the combined result of sequence specific regulatory elements, active transcription and precise developmental control. |
| Keyword | Drosophila; chorion amplification |
| 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-m3153 |
| Rights | Chen, Nan |
| Repository name | Libraries, University of Southern California |
| Repository address | Los Angeles, California |
| Repository email | http://www.usc.edu/isd/libraries/services/ask_a_librarian/email/ |
| Filename | etd-Chen-3720 |
| Archival file | uscthesesreloadpub_Volume17/etd-Chen-3720.pdf |
Description
| Title | Page 1 |
| Full text | CHORION GENE AMPLIFICATION IN DROSOPHILA MELANOGASTER by Nan Chen ————————————————————————————— A Dissertation Presented to the FACULTY OF THE GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY (MOLECULAR BIOLOGY) August 2010 Copyright 2010 Nan Chen |
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