NASCArrays Information at The BAR

Welcome to NASCArrays information at the BAR. This page hosts meta-information from the NASCArrays service (2002-2013). This information was parsed from text files available on the NASCArrays site. NASCArrays data is on iPlant server. To download experiment data from iPlant, please click on the experiment number. To download the CEL files, please click on the ftp link.

Experiment:200
Title:GENES REGULATED BY AINTEGUMENTA
Date:2005-03-11
Description:Understanding the mechanisms involved in plant growth and development is critical to achieving increased agricultural production. Because flowers, and the fruit and seeds that develop from them, represent a major source of food, the identification of new genes involved in flower development could have enormous agricultural impact. We would like to use DNA microarrays to identify genes regulated by the transcription factor AINTEGUMENTA (ANT), which is involved in floral organ initiation and growth. ANT encodes a member of the AP2/EREBP class of transcription factors. Recently, we have used an in vitro selection procedure to identify DNA sequences bound by ANT. This will allow us to directly test whether ANT regulates target genes identified by DNA microarrays in our proposed experiment. We plan to identify ANT target genes using transgenic plants containing a steroid inducible form of ANT. The inducible ANT construct consists of full-length ANT fused at its carboxyl terminal end to the ligand binding domain of the glucocorticoid receptor. The activity of the fusion protein was tested by watering 35S::ANT-GR plants daily with the steroid dexamethasone (10mm dexamethasone in 0.1% ethanol). These plants produced flowers with larger organs that closely resembled those produced by 35S::ANT plants. No phenotypes were observed in 35S::ANT-GR mock-treated plants (watered daily with 0.1% ethanol). Inflorescences from kanamycin resistant T2 plants will be used for RNA isolation as described below. The comparison probe set will consist of RNA isolated from 35S::ANT-GR inflorescences that have been treated with dexamethasone (5mm) and cycloheximide (10mm) and RNA isolated from 35S::ANT-GR inflorescences that have been treated with cycloheximide (10mm) alone. Cycloheximide is a protein synthesis inhibitor and its inclusion should prevent the identification of more downstream genes. Inflorescences will be treated by direct pipetting of approximately 25ml of the dexamethasone/cycloheximide solution or the cycloheximide solution onto the tissue. This tissue will undergo a double induction: the first application will be followed by a second application four hours later. The tissue will be harvested approximately 6 hours after the first treatment. We plan to use two arrays for biological replicates of the same experiment.
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