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Transposon-mediated embryo lethality in Arabidopsis thaliana
Theses - Dissertation
Embryo lethality in Arabidopsis can be caused by mutations in essential genes within the genome. Through the use of transposable elements, it is possible to mutate and tag these essential genes, causing the embryo lethal phenotype to appear in immature siliques of flowering Arabidopsis plants. Transposon mutagenesis also allows for revertant sector analysis of plants which are heterozygous for the mutation, thus demonstrating that the mobile transposable element is responsible for causing the embryo lethal phenotype. In this paper, I present characterization data of 44 mutant Arabidopsis lines, 29 of which contain an inheritable embryo lethal phenotype which segregates with a single transposable element. TAIL-PCR and sequencing data reveal that in 10 of these lines the transposon inserted within an identifiable gene; the insertion sites are distributed over all 5 Arabidopsis chromosomes. Revertant sector analysis of these lines, after remobilization of the transposon with the transposase enzyme, demonstrated conclusively that, in two of the 29 lines, the transposon was responsible for the disruption of the essential gene in question. Wild-type gene function was restored after the transposable element had been remobilized, supporting the theory that the element was the cause of the embryo lethal phenotype in these particular Arabidopsis lines.
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