SGER: An ORF - Rescue Vector for Crop Genome Sequencing

Iowa State University, Ames IA

Investigators

Abstract

ABSTRACT The maize genome is thought to comprise islands of lightly methylated genes in a sea of highly methylated, repetitive DNA. Complete sequencing and assembly of the whole maize genome is currently not feasible because of the repetitive DNA. Methods are beginning to be developed that will allow the regions of the maize genome that contain the genes to be filtered out for sequencing. It is expected that once validated in maize, such methods could be used for similar sequencing projects in other crop species having complex genomes. A novel expression vector system has been developed to rescue directly open reading frames (ORFs) from genomic DNA. Preliminary data suggest that this "ORF Rescue" vector system is a potentially powerful tool for gene discovery in complex crop genomes. Experiments will be performed to validate the utility of this vector system for preparing filtered template DNA for genome sequencing. The three specific aims of the project are to: 1] evaluate the ability of the ORF Rescue vector to filter genes from maize genomic DNA; 2] evaluate the ability of the ORF Rescue vector to filter genes from maize DNA cloned into Bacterial Artificial Chromosomes (BACs); 3] determine whether novel ORFs define monocot-specific genes. This work merits funding as a Small Grant for Exploratory Research (SGER) because it represents work on an untested and novel idea that is needed urgently.