Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of Arabidopsis mutants (elo2, elo3, drl1-2 and ang4) vs the wild type Ler strain


ABSTRACT: Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics. Supplemental material is available online at www.genome.org. The sequence data have been submitted to GenBank under accession number XXXXX. The GST sequence information is accessible via the CATMA database (www.catma.org) and other Arabidopsis databases, the corresponding CATMA array, or corresponding CATMA-based transcript profiling service, are available from multiple genomics facilities and the GST clone repertoire will be distributed by the Nottingham Arabidopsis Stock Centre (nasc.nott.ac.uk).

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Paul Van Hummelen 

PROVIDER: E-MEXP-30 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications


Transcript profiling is crucial to study biological systems, and various platforms have been implemented to survey mRNAs at the genome scale. We have assessed the performance of the CATMA microarray designed for Arabidopsis (Arabidopsis thaliana) transcriptome analysis and compared it with the Agilent and Affymetrix commercial platforms. The CATMA array consists of gene-specific sequence tags of 150 to 500 bp, the Agilent (Arabidopsis 2) array of 60mer oligonucleotides, and the Affymetrix gene c  ...[more]

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