Methods for tagging and mutating Arabidopsis genes with transposons.

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The objective of this award is to develop an efficient transposon tagging system for Arabidopsis thaliana, using the maize Ac transposable element. Dr. Fedoroff and her colleague will optimize the ability to detect insertion mutations in gene whose sequence is unknown by creating transposon-containing T-DNA vectors that make it possible to (1) select plants in which the tagging transposon has excised, (2) select plants in which the transposon has reinserted, and (3) screen plants for tissue-specific activation of a transposon-borne marker gene. They will also develop methods for using transposons to identify and create mutations in genes whose sequence is known. Mutations made with transposons provide a rapid means for cloning a gene, using the element as a probe, even when nothing is known about the gene's sequence. The types of mutations that will arise by insertion of the transposon tags should facilitate analysis of genes regulation by creating promoter- reporter gene fusions in vivo. The result of this award should facilitate rapid progress of the coordinated Arabidopsis genome research project.