RescueMu protocols for maize functional genomics.

Abstract

RescueMu is a modified Mu1 transposon transformed into maize to permit mutagenesis and subsequent recovery of mutant alleles by plasmid rescue. RescueMu elements insert late in the germline as well as in terminally dividing somatic (e.g., leaf) cells. Germinal insertions may result in a mutant phenotype, and RescueMu permits recovery of 5-25 kb of transposon-flanking genomic DNA without having to construct and screen genomic DNA libraries. Late somatic insertions of RescueMu do not result in a visible phenotype, but they are instead used to construct plasmid libraries of gene-enriched maize genomic DNA to facilitate the identification and sequencing of the euchromatic portion of the maize genome. This is because maize leaves contain abundant independent RescueMu somatic insertions, and 70-90% of these insertions occur preferentially into genes and not repetitive DNA. This chapter describes detailed protocols on how to obtain, generate, and use RescueMu for maize genomics, including resources developed by the Maize Gene Discovery Project (MGDP) consortium available online at ZmDB.