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Validation of de novo Bioinformatic Predictions of Arabidopsis thaliana Cis-regulatory Elements using in planta GUS Expression Assays

The study of cis-regulatory elements (CREs) will allow for increased understanding of regulation and lead to insight regarding the mechanisms governing growth, development, health, and disease. The aim of this study was to characterize the de novo in silico predictions of Arabidopsis CREs. Eight synthetic and 30 native promoter-constructs containing an eGFP/GUS reporter protein were generated for cold, genotoxic, heat, osmotic, and salt stress; the circadian clock; ABA signaling; root and epidermis tissue. Constructs were stably transformed into A. thaliana Col-0 and the effects of the CREs were evaluated by in planta stress or tissue assays using GUS expression levels. Results reveal a novel genotoxic element that specifically directs GUS expression in rosette leaves during genotoxic stress. Results also look promising for novel epidermis and root-specific elements. Results of these assays validate the de novo prediction pipeline's ability to identify novel and known CREs related to abiotic stress.

Identiferoai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/32468
Date19 July 2012
CreatorsHiu, Shuxian
ContributorsProvart, Nicholas
Source SetsUniversity of Toronto
Languageen_ca
Detected LanguageEnglish
TypeThesis

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