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Gene isolation using Arabidopsis thaliana

Some of the major scientific goals met in the Arabidopsis thaliana programme are as follows. Approximately one third of the genome has been mapped, techniques for rapidly screening yeast artificial chromosome (YAC) libraries have been developed, and a database has been established for collating mapping and genetic data. Useful frequencies of homologous recombination in Arabidopsis have been demonstrated using the reconstruction of a hygromycin resistence gene as a selection for these rare events. These constructs are being made available for testing recombination methods and deoxyribonucleic acid (DNA) delivery systems. Important new mutations affecting abscisic acid action have been isolated, and the effect of these on gene expression, lipid and sugar metabolism, and interactions with other mutants are being investigated. An 11 kb piece of DNA has been identified, using chromosome walking, that complement abi3 mutations. With this gene identified, its sequence will help reveal the mode of action of a key growth regulator in plants. T-DNA tagged loci have been identified that affect the tissue specificity of GUS expression. Several of these effect embryo specific patterns of expression. A gene encoding the APETALATA2 product has been tagged using T-DNA and isolated. This gene, which determines floral morphology together with a small group of other homeotic genes, has been further characterised by sequencing. A walk using YACs of about 1 600kb has covered the co locus, which conditions flowering time. Several recombinants have narrowed down the region containing the gene, and complementation experiments are underway to isolate clones containing the gene. Similar walks are progressing towards the fca, fwa and fve loci. In addition, epistatic interactions between the different classes of mutants are being studied. Systems that drive high frequency excision and methods for selecting reintegration events, in transposon tagging have been developed. Large populations of plants are being screened for mutations.

Reported by

John Innes Centre
Colney
NR4 7UJ Norwich
United Kingdom
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