A user's guide to restriction enzyme-mediated integration in Dictyostelium.
Restriction enzyme-mediated integration (REMI) has been used to study a number of cellular and developmental processes in Dictyostelium discoideum. In this paper we review the basics of this powerful method of introducing random mutations in Dictyostelium. Here we discuss several mutation screens that have been devised and some of the genes that have been discovered through this approach to mutagenesis. Included in this discussion is how one goes about isolating a gene that has been disrupted by REMI, and how one confirms that this disruption is actually responsible for the observed phenotype. Finally, we describe how REMI can be used as an effective teaching tool in undergraduate cell biology laboratory courses.
Guerin
Nicholas A
NA
Department of Biology, Clark University, 950 Main Street, Worcester, MA 01610, USA.
Larochelle
Denis A
DA
D000818:Animals; D004262:DNA Restriction Enzymes; D004023:Dictyostelium / Q000235:genetics*; D005800:Genes, Fungal; D017125:Genes, Protozoan; D008957:Models, Genetic; D016296:Mutagenesis; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics; D015183:Restriction Mapping / Q000379:methods*
D015800:Protozoan Proteins; D004262:DNA Restriction Enzymes