Two large-insert soybean genomic libraries constructed in a binary vector: applications in chromosome walking and genome wide physical mapping (original) (raw)

Abstract

Large DNA insert libraries in binary T-DNA vectors can assist in the isolation of the gene(s) underlying a quantitative trait locus (QTL). Binary vectors facilitate the transfer of large-insert DNA fragments containing a QTL from E. coli to Agrobacterium sp. and then to plants. We constructed two soybean large-insert libraries from cv. Forrest in the pCLD04541 (V41) binary vector after partial digestion of genomic high-molecular-weight DNA with _Bam_HI or _Hin_dIII. The libraries contain 76,800 clones with an average insert size of 125 kb, and therefore represent 9.5-fold haploid genome equivalents. Colony hybridization using a chloroplast-specific probe infers that the libraries contain less than 0.5% clones of chloroplast DNA origin. These two libraries have provided clones for physical mapping of the soybean genome and for isolation of a number of disease resistance genes. One microsatellite marker was identified from the clone that hybridized to the Bng122 RFLP probe. The sequence-tagged site was used for genetic mapping and marker-assisted selection for genes underlying resistance to the soybean cyst nematode and sudden death syndrome.

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Authors and Affiliations

  1. Department of Plant Soil and General Agriculture, Southern Illinois University at Carbondale, Room 176. Carbondale, IL 62901–4415, USA Fax: +618 453 7457 e-mail: http://www.coalab.siu.edu/genome/meksemk@siu.edu, , , , , , US
    K. Meksem, K. Zobrist, E. Ruben, D. Hyten & D. A. Lightfoot
  2. Department of Soil and Crop Sciences, Crop Biotechnology Center, Texas A & M University, College Station, TX 77843, USA, , , , , , US
    T. Quanzhou & H-B. Zhang

Authors

  1. K. Meksem
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  2. K. Zobrist
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  3. E. Ruben
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  4. D. Hyten
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  5. T. Quanzhou
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  6. H-B. Zhang
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  7. D. A. Lightfoot
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Received: 7 May 1999 / Accepted: 18 February 2000

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Meksem, K., Zobrist, K., Ruben, E. et al. Two large-insert soybean genomic libraries constructed in a binary vector: applications in chromosome walking and genome wide physical mapping.Theor Appl Genet 101, 747–755 (2000). https://doi.org/10.1007/s001220051540

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