title: Rapid recombination mapping for high-throughput genetic screens in Drosophila.
abstract: Mutagenesis screens are a staple of classical genetics. Chemical-induced mutations, however, are often difficult and time-consuming to identify. Here, we report that recombination analysis with pairs of dominant visible markers provides a rapid and reliable strategy to map mutations in Drosophila melanogaster. This method requires only two generations and a total of six crosses in vials to estimate the genetic map position of the responsible lesion with high accuracy. This genetic map position can then be reliably used to identify the mutated gene through complementation testing with an average of nine deficiencies and Sanger sequencing. We have used this approach to successfully map a collection of mutations from an ethyl methanesulfonate-based mutagenesis screen on the third chromosome. We propose that this method also may be used in conjunction with whole-genome sequencing, particularly when multiple independent alleles of the mutated locus are not available. By facilitating the rapid identification of mutated genes, our mapping strategy removes a primary obstacle to the widespread use of powerful chemical mutagenesis screens to understand fundamental biological phenomena.
authors.affiliation: Division of Pharmaceutical Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53705-2222.
authors.forename: Anne L
authors.initials: AL
authors.lastname: Sapiro
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Ihry
authors.forename: Derek L
authors.initials: DL
authors.lastname: Buhr
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Konieczko
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Ives
authors.forename: Anna K
authors.initials: AK
authors.lastname: Engstrom
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Wleklinski
authors.forename: Kristin J
authors.initials: KJ
authors.lastname: Kopish
authors.forename: Arash
authors.initials: A
authors.lastname: Bashirullah
mesh_terms: D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D004330:Drosophila / Q000187:drug effects / Q000235:genetics; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D005799:Genes, Dominant; D005819:Genetic Markers; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D016296:Mutagenesis / Q000187:drug effects; D009154:Mutation; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics*; D012469:Salivary Glands / Q000187:drug effects / Q000502:physiology
chemical_list: D005819:Genetic Markers; D005020:Ethyl Methanesulfonate
keywords: EMS mutagenesis; dominant markers; persistent salivary glands (PSG)