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Originally published as Genetics Published Articles Ahead of Print on January 21, 2007.
Genetics, Vol. 175, 1047-1058, March 2007, Copyright © 2007
doi:10.1534/genetics.106.065573
Single-Gene Detection and Karyotyping Using Small-Target Fluorescence in Situ Hybridization on Maize Somatic Chromosomes
Jonathan C. Lamb*,1,
Tatiana Danilova*,1,
Matthew J. Bauer*,
Julie M. Meyer
,
Jennifer J. Holland*,
Michael D. Jensen
and
James A. Birchler*,2
* Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211, Department of Agriculture, Truman State University, Kirksville, Missouri 63501 and Department of Biochemistry, University of Missouri, Columbia, Missouri 65211
2 Corresponding author: 117 Tucker Hall, University of Missouri, Columbia, MO 65211.
E-mail: birchlerj{at}missouri.edu
Combined with a system for identifying each of the chromosomes in a genome, visualizing the location of individual genetic loci by fluorescence in situ hybridization (FISH) would aid in assembling physical and genetic maps. Previously, large genomic clones have been successfully used as FISH probes onto somatic chromosomes but this approach is complicated in species with abundant repetitive elements. In this study, repeat-free portions of sequences that were anchored to particular chromosomes including genes, gene clusters, large cDNAs, and portions of BACs obtained from public databases were used to label the corresponding physical location using FISH. A collection of probes that includes at least one marker on each chromosome in the maize complement was assembled, allowing a small-target karyotyping system to be developed. This set provides the foundation onto which additional loci could be added to strengthen further the ability to perform chromosomal identification in maize and its relatives. The probes were demonstrated to produce signals in several wild relatives of maize, including Zea luxurians, Z. diploperennis, and Tripsacum dactyloides.
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