title: Regional mutagenesis using Dissociation in maize.
abstract: We describe genetic screens, molecular methods and web resources newly available to utilize Dissociation (Ds) as an insertional mutagen in maize. Over 1700 Ds elements have been distributed throughout the maize genome to serve as donor elements for local or regional mutagenesis. Two genetic screens are described to identify Ds insertions in genes-of-interest (goi). In scheme I, Ds is used to generate insertion alleles when a recessive reference allele is available. A Ds insertion will enable the cloning of the target gene and can be used to create an allelic series. In scheme II, Ds insertions in a goi are identified using a PCR-based screen to identify the rare insertion alleles among a population of testcross progeny. We detail an inverse PCR protocol to rapidly amplify sequences flanking Ds insertion alleles and describe a high-throughput 96-well plate-based DNA extraction method for the recovery of high-quality genomic DNA from seedling tissues. We also describe several web-based tools for browsing, searching and accessing the genetic materials described. The development of these Ds insertion lines promises to greatly accelerate functional genomics studies in maize.
authors.affiliation: Boyce Thompson Institute for Plant Research, Ithaca, NY 14853, USA.
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Ahern
authors.forename: Prasit
authors.initials: P
authors.lastname: Deewatthanawong
authors.forename: Justin
authors.initials: J
authors.lastname: Schares
authors.forename: Michael
authors.initials: M
authors.lastname: Muszynski
authors.forename: Rebecca
authors.initials: R
authors.lastname: Weeks
authors.forename: Erik
authors.initials: E
authors.lastname: Vollbrecht
authors.forename: Jon
authors.initials: J
authors.lastname: Duvick
authors.forename: Volker P
authors.initials: VP
authors.lastname: Brendel
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Brutnell
mesh_terms: D019295:Computational Biology; D004251:DNA Transposable Elements / Q000235:genetics*; D020407:Internet; D016254:Mutagenesis, Insertional / Q000379:methods*; D012984:Software; D003313:Zea mays / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements