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Nucleic Acids Research, 1990, Vol. 18, No. 11 3271-3279
© 1990

GENOME STRUCTURE AND MAPPING

Amplification of genomic sequences flanking transposable elements in host and heterologous plants: a tool for transposon tagging and genome characterization

David J. Earp1, Brenda Lowe1 and Barbara Baker1,2,*

1Plant Gene Expression center, University of California Berkley/USDA 800, Buchanan St., Albany, CA 94710 2Department of Plant Pathology, UNiversity of Calfornia Berkely, CA 94710, USA

*To whom correspondence should be addressed

Received February 21, 1990. Accepted April 30, 1990.

The isolation of sequences flanking integrated transposable elements is an important step in gene tagging strategies. We have demonstrated that sequences flanking transposons integrated Into complex genomes can be simply and rapidly obtained using the polymerase chain reaction. Amplification of such sequences was established in a model system, a transgenic tobacco plant carrying a single Ac element, and successfully applied to the cloning of a specific Spm element from a maize line carrying multiple Spm hybridizing sequences. The described utilization of methylation sensitive restriction enzymes (including those with degenerate recognition sequences) in the generation of templates for amplification will simplify the cloning and mapping of genomic sequences adjacent to transposable elements.


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