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Transposon Display identifies individual transposable elements in high copy number lines
Authors:Van den Broeck D  Maes T  Sauer M  Zethof J  De Keukeleire P  D'hauw M  Van Montagu M  Gerats T
Institution:Laboratorium voor Genetica, Universiteit Gent, K.L.Ledeganckstraat 37, B-9000 Gent, Belgium.
Abstract:The dTph1 transposable element family of Petunia hybrida line W138 consists of between 100 and 200 members. A strategy that allows simultaneous detection of individual elements is described. Sequences flanking dTph1 elements are amplified by means of a ligation-mediated PCR. The resulting fragments are locus-specific and can be analysed by polyacrylamide gel electrophoresis. One of the applications of Transposon Display is the isolation of dTph1-tagged genes. Fragments that co-segregate with a mutant phenotype can be extracted from the gel and reamplified, providing access to tagged genes, as demonstrated in a reconstruction experiment. Data on the molecular identification of a phenotypic mutant, isolated in a random tagging experiment is also presented. Upon sequencing, the obtained candidate fragment was found to be identical to part of the previously identified Fbp1 gene.
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