Plant & Animal Genome IX Conference Plant & Animal Genome IX Conference
Poster: Large Insert Libraries, Gene Isolation, Etc.
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Positional cloning of the interesting genes has been made more feasible by the development of detailed genetical and physical maps of major crops, such as wheat and barley, identification of major QTLs responsible for agronomically important traits, and the development of large insert libraries. However, large genome size, the presence of multiple repetitive elements, as well as a limited amount of colinearity between model species and major cereal crops makes the task of chromosome walking extremely difficult. One of the ways to alleviate the problem of plant genomes' low information content is to concentrate on the portions of the genome which are meaningful. We utilized restriction endonuclease McrBC to separate heavily methylated and undermethylated fractions of genomic DNA. This approach permitted us to generate a library from undermethylated, gene-enriched fragments of barley genome. Such an approach also circumvented potential problems associated with gene isolation through the characterization of EST libraries. The range of these problems includes difficulties in cloning of the rare messages, unaccounting for the regulatory gene sequences and skewed representation of multigene family members. Targeted isolation and cloning of the selected gene-rich regions might provide great benefits for characterization of the interesting genome areas, while greatly reducing complexity of the task.