Transposon Display identifies individual transposable elements in high copy number lines.
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.
Van den Broeck
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Laboratorium voor Genetica, Universiteit Gent, K.L.Ledeganckstraat 37, B-9000 Gent, Belgium.
Maes
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Sauer
M
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Zethof
J
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De Keukeleire
P
P
D'hauw
M
M
Van Montagu
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M
Gerats
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