|Chen, Zengjian - UNIVERSITY OF MINNESOTA|
|Phillips, Ronald - UNIVERSITY OF MINNESOTA|
Submitted to: Agronomy Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: November 2, 1995
Publication Date: N/A
Technical Abstract: A genomic subtraction technique was used to selectively isolate maize (Zea mays L.) sequences from a maize chromosome 3 addition line of oat (Avena sativa L.). Two representative DNA samples, one from the addition line and one from the oat parent, were prepared using BamHI digestion followed by PCR amplification. Subtractive hybridization with an excess of oat DNA then was used to remove common DNA fragments present in both samples. The remaining DNA fragments were cloned into a plasmid vector. A DNA library containing 400 clones was constructured by this method. In a test of 18 clones, two (11%) detected maize-specific repetitive DNA sequences and seven (39%) showed strong hybridization to genomic DNAs of maize but weak hybridization to oat. Among these latter seven clones, two of them detected DNA sequences specific to chromosome 3 of maize, the chromosome retained in the original maize addition line of oat. The other five detected repetitive sequences on most or all of the other maize chromosomes. We estimate that the maize DNA sequences were enriched approximately 10-20 fold by this genomic subtraction technique; however, most of the isolated DNA fragments detected multiple or repeated DNA sequences.