Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Activator ( Ac )

The bronze ( bz ) locus of maize has been cloned by an indirect procedure utilizing the cloned transposable controlling element Activator ( Ac ). Restriction endonuclease fragments of maize DNA were cloned in bacteriophage λ and recombinant phage with homology to the center of the Ac element were isolated. The cloned fragments were analyzed to determine which contained sequences that were structurally identical to a previously isolated Ac element. Two such fragments were identified. Sequences flanking the Ac element were subcloned and used to probe genomic DNA from plants with well-defined mutations at the bz locus. By this means, it was established that one of the genomic clones contained a bz locus sequence. The subcloned probe fragment was then used to clone a nonmutant Bz allele of the locus. The method described here should prove useful in cloning other loci with Ac insertion mutations.