PI, Joseph R. Ecker, The Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, Email: ecker@salk.edu Co-PI, Ronald W. Davis, Stanford Genome Technology Center, Stanford University, 855 California Ave, Palo Alto, CA 94304, Email: dbowe@sequence.stanford.edu Co-PI, Athanasios Theologis, Plant Gene Expression Center, U.C. Berkeley, 800 Buchanan Street, Albany, CA 94022, Email theo@nature.berkeley.edu 1. Project Summary In order to carry out functional genomic and proteomic studies using the recently completed Arabidopsis genomic sequence, we must be able to readily manipulate and express each of the genes as proteins. Unfortunately, current computational approaches for Arabidopsis gene prediction are unable to precisely predict or, in some cases, even recognize many of the genes. These limitations prohibit the use of new emerging technologies for global gene functional analysis. The aim of our program is to experimentally determine the transcription units for all Arabidopsis genes and create a gold standard set of open-reading-frame (ORF) clones. This will provide an accurate determination of each of the gene structures and allow for the construction of full-length cDNAs for each gene. Determining the full-length sequences for the transcription units will resolve ambiguities in the computationally annotated genomic sequence, allowing precise position of introns/exons and 5' transcription start and 3' polyadenyation addition sites,. The identification of full-length cDNAs for all Arabidopsis genes is of primary importance for the entire plant biology community as these clones will be essential for many future functional genomic and proteomic studies. 2. Deliverables of the SSP Consortium. Isolation and complete sequencing of full-length cDNAs for 8,000 genes with immediate deposit of cDNA sequences in GenBank. SSP-sequenced RAFL cDNAs are available through the RIKEN Bioresource Center Construction of 8,000 gold standard ORF clones in a universal recombination plasmid vector (pUNI). The ORF clones are fully sequence validated and are error-free. These clones are deposited in the Arabidopsis Biological Resource Center (ABRC) at Ohio State University with no material transfer agreement. Among the 8,000 ORF clones, 7,000 will be constructed by PCR from full-length cDNAs and 1,000 will be identified from non-expressed annotated (hypothetical) genes. Identification of novel Arabidopsis transcription units using custom Affymetrix genome tiling arrays and mRNA samples prepared from various plant tissues and conditions. URL for SSP Consortium Project Deliverables: http://signal.salk.edu/SSP/index.html
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