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Characterization and application of soybean YACs to molecular cytogenetics

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Abstract

Yeast artificial chromosomes (YACs) are widely used in the physical analysis of complex genomes. In addition to their value in chromosome walking for map-based cloning, YACs represent excellent probes for chromosome mapping using fluorescence in situ hybridization (FISH). We have screened such a library for low-copy-number clones by hybridization to total genomic DNA. Four clones were chosen for chromosome tagging based upon their low or moderate signal. By using degenerate oligonucleotide-primed PCR (DOP-PCR), we were able to use relatively small amounts of soybean YAC DNA, isolated directly by preparative pulsed-field gel electrophoresis, as FISH probes for both metaphase chromosome spreads and interphase nuclei. FISH chromosomal analysis using the three of the clones as probes resulted in relatively simple hybridization patterns consistent with a single homologous locus or two homoeologous loci. The fourth YAC probe resulted in a diffuse hybridization pattern with signal on all metaphase chromosomes. We conclude that YACs represent a valuable source of probes for chromosomal analysis in soybean.

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Author notes

  1. T. Zhu & L. Shi

    Present address: Section of Plant Biology, University of California, 95616, Davis, CA, USA

  2. R. P. Funke

    Present address: The Beadle Center, The University of Nebraska-Lincoln, 68583, Lincoln, NE, USA

Authors and Affiliations

  1. Department of Biological Sciences, Northern Arizona University, 86011-5640, Flagstaff, AZ, USA

    T. Zhu, L. Shi & P. Keim

  2. Plant Molecular Genetics and Center for Legume Research, University of Tennessee, 37901-1071, Knoxville, TN, USA

    R. P. Funke & P. M. Gresshoff

Authors
  1. T. Zhu
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  2. L. Shi
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  3. P. Keim
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  4. R. P. Funke
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  5. P. M. Gresshoff
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Communicated by J. Schell

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Zhu, T., Shi, L., Keim, P. et al. Characterization and application of soybean YACs to molecular cytogenetics. Molec. Gen. Genet. 252, 483–488 (1996). https://doi.org/10.1007/BF02173014

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  • DOI: https://doi.org/10.1007/BF02173014

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