Plant Physiology Preview
Published on December 5, 2002; 10.1104/pp.013474
Received August 23, 2002
Returned for revision October 2, 2002
Accepted October 8, 2002
Characterization of Three Maize Bacterial Artificial Chromosome Libraries toward Anchoring of the Physical Map to the Genetic Map Using High-Density Bacterial Artificial Chromosome Filter Hybridization
Young-Sun Yim , Georgia L. Davis *, Ngozi A. Duru , Theresa A. Musket , Eric W. Linton , Joachim W. Messing , Michael D. McMullen , Carol A. Soderlund , Mary L. Polacco , Jack M. Gardiner , and Edward H. Coe Jr.
Department of Agronomy, University of Missouri, 1-87 Agriculture, Columbia, Missouri 65211 (Y.-S.Y., G.L.D., N.A.D., T.A.M., M.D.M., M.L.P., J.M.G., E.H.C.); Waksman Institute, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854 (E.W.L., J.W.M.); United States Department of Agriculture-Agricultural Research Service, Plant Genetics Research Unit, 210 Curtis Hall, Columbia, Missouri 65211 (M.D.M., M.L.P., E.H.C.); and Plant Science Department, University of Arizona, Tucson, Arizona 85721 (C.A.S.)
* Corresponding author; email: DavisGe{at}missouri.edu.
Three maize (Zea mays) bacterial artificial chromosome (BAC) libraries were constructed from inbred line B73. High-density filter sets from all three libraries, made using different restriction enzymes (HindIII, EcoRI, and MboI, respectively), were evaluated with a set of complex probes including the185-bp knob repeat, ribosomal DNA, two telomere-associated repeat sequences, four centromere repeats, the mitochondrial genome, a multifragment chloroplast DNA probe, and bacteriophage  . The results indicate that the libraries are of high quality with low contamination by organellar and -sequences. The use of libraries from multiple enzymes increased the chance of recovering each region of the genome. Ninety maize restriction fragment-length polymorphism core markers were hybridized to filters of the HindIII library, representing 6x coverage of the genome, to initiate development of a framework for anchoring BAC contigs to the intermated B73 x Mo17 genetic map and to mark the bin boundaries on the physical map. All of the clones used as hybridization probes detected at least three BACs. Twenty-two single-copy number core markers identified an average of 7.4 ± 3.3 positive clones, consistent with the expectation of six clones. This information is integrated into fingerprinting data generated by the Arizona Genomics Institute to assemble the BAC contigs using fingerprint contig and contributed to the process of physical map construction.
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