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Prospects of applying a combination of DNA transposition and site-specific recombination in plants: a strategy for gene identification and cloning
Authors:Mark J. J. van Haaren  David W. Ow
Affiliation:(1) Department of Genetics, Institute for Molecular Biological Sciences, Biocentrum Amsterdam, De Boelelaan 1087, 1081 HV Amsterdam, Netherlands;(2) Plant Gene Expression Center, U.S., Department of Agriculture, 800 Buchanan Street, 94710 Albany, CA, USA
Abstract:The concept of gene identification and cloning using insertional mutagenesis is well established. Many genes have been isolated using T-DNA transformation or transposable elements. Maize transposable elements have been introduced into heterologous plant species for tagging experiments. The behaviour of these elements in heterologous hosts shows many similarities with transposon behaviour in Zea mays. Site-specific recombination systems from lower organisms have also been shown to function efficiently in plant cells. Combining transposon and site-specific recombination systems in plants would create the possibility to induce chromosomal deletions. This lsquotransposition-deletionrsquo system could allow the screening of large segments of the genome for interesting genes and may also permit the cloning of the DNA corresponding to the deleted material by the same site-specific recombination reaction in vitro. This methodology may provide a unique means to construct libraries of large DNA clones derived from defined parts of the genome, the phenotypic contribution of which is displayed by the mutant carrying the deletion.
Keywords:transposition  Ac/Ds  site-specific recombination  Cre/lox  chromosomal rearrangements  deletion mutagenesis
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