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Post-graduate theses

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Identifier

000334431

Title

Χαρακτηρισμός της δομής και δράσης βασικών υποκινητών του αμφίποδου καρκινοειδούς Parhyale hawaiensis

Alternative Title

Characterization of core promoters structure and function in the amphipod crystacean Parhyale hawaiensis

Author

Κωνσταντινίδης, Νικόλαος

Thesis advisor

Δελιδάκης, Χρήστος

Abstract

The advancement of new model organisms is necessary for the comparison of information derived from already established ones (D. melanogaster, C. elegans, mouse) and for the acquisition of information concerning the evolution of these species. As a prerequisite, genetic tools for the manipulation of these organisms should be developed. The primary aim of this thesis is the characterization of the structure and function of two basal promoters from the emerging model crustacean Parhyale hawaiensis, for their future utilization in transformation experiments. We used 5’ RACE to map the basal promoters of the PhHS and PhMS elements of the hsp70 gene family. We identified the position of the transcription start site, the TATA boxes, splice acceptors and splice donors relative to the HSF and bHLH binding sites positions. We then prepared and injected one cell-stage embryos with two plasmid constructs; these constructs carried, between the ends of the Minos transposable element, a marker gene (DsRed or EGFP) under the transcriptional control of the 3xP3 regulatory element in conjunction with the Drosophila hsp70 basal promoter and another marker gene under the control of the 3xP3 or DC5 regulatory elements in conjunction with the Parhyale hsp70 basal promoter. After analysis of first generation embryos (G1), we came to the conclusion that the Drosophila basal promoter is not as functional as the endogenous one when used in Parhyale. We also compared the activation patterns of 3xP3 and DC5 regulatory elements in Parhyale hawaiensis in order to determine their utility as markers of stable transformation.

Physical description

48 σ. : εικ. ; 30 εκ.

Language

Greek

Subject

Drosophila

PhHS

PhMS

Issue date

2008-12-04

Collection

School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses