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.

[1]  M. Van Montagu,et al.  Insertion mutagenesis and study of transposable elements using a new unstable virescent seedling allele for isolation of haploid petunia lines. , 1996, The Plant Journal.

[2]  G. Bernardi,et al.  Compositional bimodality of the nuclear genome of tobacco. , 1991, Nucleic acids research.

[3]  H. Huits,et al.  Molecular characterization of a nonautonomous transposable element (dTph1) of petunia. , 1990, The Plant cell.

[4]  W. Dawson,et al.  HOST-RANGE DETERMINANTS OF PLANT VIRUSES , 1992 .

[5]  H. Huits,et al.  Gene tagging in Petunia hybrida using homologous and heterologous transposable elements , 1989 .

[6]  Yongbiao Xue,et al.  Cloning and structural analysis of the anthocyanin pigmentation locus Rt of Petunia hybrida: characterization of insertion sequences in two mutant alleles. , 1994, The Plant journal : for cell and molecular biology.

[7]  R. Koes,et al.  A general method to isolate genes tagged by a high copy number transposable element. , 1995, The Plant journal : for cell and molecular biology.

[8]  P. Vos,et al.  AFLP: a new technique for DNA fingerprinting. , 1995, Nucleic acids research.

[9]  J. Salinas,et al.  Non-random distribution of transposable elements in the nuclear genome of plants. , 1993, Nucleic Acids Research.

[10]  G. Angenent,et al.  Petal and stamen formation in petunia is regulated by the homeotic gene fbp1. , 1993, The Plant journal : for cell and molecular biology.

[11]  V. Walbot.  Strategies for Mutagenesis and Gene Cloning Using Transposon Tagging and T-DNA Insertional Mutagenesis , 1992 .

[12]  L. Mur,et al.  Targeted gene inactivation in petunia by PCR-based selection of transposon insertion mutants. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[13]  J. Mol,et al.  The No Apical Meristem Gene of Petunia Is Required for Pattern Formation in Embryos and Flowers and Is Expressed at Meristem and Primordia Boundaries , 1996, Cell.

[14]  E. Coen,et al.  A common gene regulates pigmentation pattern in diverse plant species , 1992, Cell.

[15]  M. Freeling,et al.  Genetic characterization of the Mutator system in maize: behavior and regulation of Mu transposons in a minimal line. , 1995, Genetics.

[16]  E. Coen,et al.  floricaula: A homeotic gene required for flower development in antirrhinum majus , 1990, Cell.