Amplification of genomic sequences flanking transposable elements in host and heterologous plants: a tool for transposon tagging and genome characterization.
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.
Earp
D J
DJ
Plant Gene Expression Center, University of California, Albany 94710.
Lowe
B
B
Baker
B
B
D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D005784:Gene Amplification; D017343:Genes, Plant; D008745:Methylation; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D013698:Templates, Genetic; D014026:Nicotiana / Q000235:genetics*; D003313:Zea mays / Q000235:genetics
D004251:DNA Transposable Elements