A Sequence-Indexed Library of Insertion Mutations in the Arabidopsis Genome
With the availability of the entire Arabidopsis genome sequence, one
of the next challenges is to uncover the functions of the more than
25,000 genes in this reference plant. Given the scope of the NSF 2010
program, -to identify the function of all Arabidopsis genes in the
next decade-, an efficient and cost effective approach is necessary
to identify mutations in all genes. The goal of this program is to
create a sequence-indexed library of mutations in the Arabidopsis
genome. The Salk Institute Genome Analysis Laboratory
(SIGnAL) will use high-throughput genome sequencing
methods to identify the sites of insertion of Agrobacterium T-DNA in
the Arabidopsis genome. T-DNA transformed plants from the
Alonso/Crosby/Ecker collection will be grown, genomic DNA will be
prepared, T-DNA flanking plant DNA will be recovered and sequenced.
Insertion site sequences will be aligned with the Arabidopsis genome
sequence and gene annotation will be added. The data will be made
available via a web accessible graphical
interface-T-DNAExpress- (http://signal.salk.edu/cgi-bin/tdnaexpress)
that will provide both text and DNA searches of the insertion
sequence database. All DNA sequences will be deposited into GenBank
and also provided to The Arabidopsis
Information Resource (TAIR)
Seeds from the T-DNA insertion lines will be deposited with the
Arabidopsis Biological Resource Center (ABRC) at Ohio State University
The ABRC will propagate and will distribute seeds to the community.
The creation of a searchable database containing the insertion site
information and the availability of the corresponding mutant lines in
public stock centers will provide researchers with ready access to
mutants in their genes of interest, allowing the testing of
hypotheses about gene function at an unprecedented rate.
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