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United States Department of Agriculture

Agricultural Research Service

Title: Using Restriction Enzymes As a Method of Genome Filtering in Wheat

item Fellers, John

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: June 7, 2005
Publication Date: January 15, 2006
Citation: Fellers, J.P. 2006. Using restriction enzymes as a method of genome filtering in wheat. Plant and Animal Genome Conference Abstract.

Technical Abstract: Bread wheat (Triticum aestivum) contains one of the largest genomes of the plant kingdom and is a significant challenge when considered for genome sequencing. Wheat is known to have gene-rich regions within the genome interspersed by vast amounts of repetitive DNA. It is understood that a filtering technique would be of significant benefit when targeting gene-containing regions for sequencing. Previous work has indicated that the restriction enzyme, Pst1 can increase the presence of low copy DNA when used to digest genomic DNA. To evaluate this on a larger scale, genomic DNA of the bread wheat 'Chinese Spring' was digested with the methyl sensitive restriction enzymes, Sac1, Sal1, Pst1, and AatII. DNA fragments of >400 bp were subcloned into cloning vector, isolated, and single pass sequenced. Preliminary evidence indicates that this technique can enrich by more than 35%, depending on the enzyme used. The sequencing results will be evaluated for the presence of open reading frames, repetitive DNA and low copy DNA. The results will be reported here.

Last Modified: 4/22/2015
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