Fluorescence in situ hybridization to localize transgenes in plant chromosomes.

Abstract

Production of transgenic plants is now routine for many of our crop species. Methods for the detailed molecular analysis of transgenic plants are available, but often the exact location of the transgene within the crop genome is poorly understood. As a starting point to understanding more about the site of transgene insertion, transgenes can be physically located using fluorescence in situ hybridization (FISH). This technique allows transgenes to be located to specific chromosome regions following the hybridization of a fluorescent labelled probe to a chromosome spread. The technique is sensitive enough to detect single transgene copies and can reveal information about the complexity of a transgene insertion site as well as identifying plants homozygous for the transgene. A FISH method is described that has been used successfully to detect single-transgene copies in mitotic metaphase chromosome preparations of wheat and barley.