3.
RescueMu Protocols for Maize Functional Genomics
| Abstract |
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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.
Affiliation(s): (2) Laboratory of Crop Genomics, Department of Plant Agriculture, University of Guelph, Ontario, Canada
Book Title: Plant Functional Genomics
Series: Methods in Molecular Biology | Volume: 236 | Pub. Date: Aug-06-2003 | Page Range: 37-58 | DOI: 10.1385/1-59259-413-1:37
Subject: Plant Sciences
Key Words: Mutator - RescueMu - maize - genomics - transposon - genome survey sequence - plasmid rescue - techniques
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