Poster: Late and Moved Abstracts
Abs #
1402: A New Reverse-Genetic Resource for Knocking-Out Small Genes and Tandemly
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Presenter: |
Krysan, Patrick J, fpat@biotech.wisc.edu |
Authors | Krysan, Patrick J (A) Woody, Scott (A) Monson, Sean (A) Jester, Peter (A) Amasino, Richard (A) Sussman, Michael (A) | | Affiliations: |
(A): University of Wisconsin
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It has become apparent that reverse-genetic analysis is a powerful method for analyzing gene function in Arabidopsis. Several resources are currently available that make knockout mutants of Arabidopsis genes easily available to researchers throughout the world. Because these systems are all based on random mutagenesis, the principles of probability dictate that null alleles for many small genes will not be available in the current collections. In addition, if you are interested in a family of tandemly duplicated genes, there is currently no simple system for creating a plant that carries knockout alleles for multiple members of a tandem gene array.
We are currently in the process of developing a new reverse-genetic resource that should provide the means to obtain knockout alleles for genes that have escaped mutagenesis in the currently available reverse-genetic collections. In addition, the system also provides the means to knockout multiple members of a tandemly duplicated set of genes. The resource is based on a new collection of T-DNA lines that were constructed using a new T-DNA vector that contains a DS transposon and LoxP sites. We are using TAIL PCR to map the locations of thousands of individual T-DNA inserts from this collection. The DS transposon in this vector can be mobilized by crossing the T-DNA line to a source of transposase. Because DS transposition is biased towards tightly linked regions of the genome, it should be possible to saturate a local area of the genome with transposed DS elements, thereby generating a knockout of the small gene of interest. In addition, Cre/Lox recombination can also be used to create small chromosomal deletions that remove multiple members of a tandem gene array.
