Fluorescent transgenes to study interphase chromosomes in living plants

Abstract

Fluorescence tagging of genomic sites through the use of bacterial operator/repressor systems combined with fluorescent proteins permits high-resolution analysis of interphase chromosomes in living cells. This technique has been used to study interphase chromosome arrangement and dynamics in yeast, Drosophila, and mammalian cells, but is only beginning to be exploited in plant systems. In this chapter, we describe methods for producing and identifying Arabidopsis thaliana plants harbouring fluorescence-tagged transgenes. The use of these plants to analyze various aspects of interphase chromosome organization and dynamics in living cells using 3D wide-field fluorescence microscopy is discussed. Potential problems encountered when utilizing this technology in plants are considered. © 2008 Humana Press.