A bacterial artificial chromosome library for soybean and identification of clones near a major cyst nematode resistance gene |
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| Authors: | D. Danesh S. Peñuela J. Mudge R. L. Denny H. Nordstrom J. P. Martinez N. D. Young |
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| Affiliation: | (1) Department of Plant Pathology, 495 Borlaug Hall, 1991 Upper Buford Circle, University of Minnesota, St. Paul, Minnesota 55108, USA Fax: 612-625-9728 E-mail: neviny@tc.umn.edu, US;(2) Department of Agronomy and Plant Genetics, 411 Borlaug Hall, 1991 Upper Buford Circle, University of Minnesota, St. Paul, Minnesota 55108, USA, US |
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| Abstract: | We constructed a bacterial artificial chromosome (BAC) library for soybean (Glycine max) consisting of approximately 30 000 clones with an average insert size of 120 kilobase pairs. The library was successfully screened with restriction fragment length polymorphism (RFLP) and microsatellite markers tightly linked to a major resistance gene for the cyst nematode, Heterodera glycines. Since many soybean RFLPs hybridize to duplicate loci, BACs homologous to duplicate RFLP loci were distinguished by digestion with the restriction enzyme originally used to map the RFLP, followed by a comparison of the hybridizing fragments. Linkage mapping of BAC clones identified with markers linked to the cyst nematode resistance gene demonstrated that these clones were located at the expected chromosomal positions and that there were no indications of chimeras within the genomic inserts. Received: 3 July 1997/Accepted: 26 August 1997 |
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| Keywords: | Chromosome walking Gene mapping Glycine max Heterodera glycines High-molecular-weight DNA Positional cloning |
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