First published online December 5, 2002; 10.1104/pp.013474
Plant Physiol, December 2002, Vol. 130, pp. 1686-1696
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
Hybridization1
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.)
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 6×
coverage of the genome, to initiate development of a framework for
anchoring BAC contigs to the intermated B73 × 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.
1
This work was supported by the National Science
Foundation (Plant Genome grant nos. DBI 9872655 and 9975618).
*
Corresponding author; e-mail DavisGe{at}missouri.edu;
fax 573-882-1469.
© 2002 American Society of Plant Biologists
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