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Construction of a BAC Library for a Defoliating Insect-Resistant Soybean and Identification of Candidate Clones Using a Novel Approach

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Abstract

Positional cloning of an insect-resistance quantitative trait locus (QTL) requires the construction of a large-insert genomic DNA library from insect-resistant genotypes. To facilitate cloning of a major defoliating insect-resistance QTL on linkage group M of the soybean genetic map, a bacterial artificial chromosome (BAC) library for PI 229358 was constructed and characterized. The HindIII BAC library contains 55,296 clones with an average insert size 131 kb. This library represents a 6-fold soybean haploid genome equivalents, allowing a 99.8% probability of recovering any specific sequence of interest in soybean. BAC filters were screened with a genomic DNA probe Sat_258sc2 obtained through genome walking from flanking sequences of a simple sequence repeat (SSR) marker, Sat_258, which links to the insect-resistance QTL. Thirteen BAC clones were identified positive for Sat_258sc2, and two of them were confirmed to carry Sat_258. The results suggest that this library is useful in positional cloning of the major insect-resistance QTL, and the approach presented here can be used to screen a BAC library for a SSR marker without requiring the creation of BAC pools.

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Acknowledgments

We would like to thank Michael Atkins for his assistance with producing BAC filters. This research was supported by the USDA National Research Initiative Competitive Grants Program (grant no. 2002-01460) and by state and federal funding allocated to the Georgia Agricultural Experiment Stations.

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Author notes

  1. S. Zhu

    Present address: Monsanto Company, GG6A, 700 Chesterfield Pkwy West, Chesterfield, MO, 63017, USA

Authors and Affiliations

  1. Center for Applied Genetic Technologies and Department of Crop & Soil Sciences, University of Georgia, Athens, GA, 30602, USA

    S. Zhu, H. R. Boerma & W. A. Parrott

  2. Clemson University Genomics Institute, Clemson University, Clemson, SC, 29634, USA

    C. A. Saski & J. P. Tomkins

  3. Department of Entomology, University of Georgia, Athens, GA, 30602, USA

    J. N. All

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  1. S. Zhu
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  2. C. A. Saski
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  4. J. P. Tomkins
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  5. J. N. All
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  6. W. A. Parrott
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Correspondence to W. A. Parrott.

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Zhu, S., Saski, C.A., Boerma, H.R. et al. Construction of a BAC Library for a Defoliating Insect-Resistant Soybean and Identification of Candidate Clones Using a Novel Approach. Plant Mol Biol Rep 27, 229–235 (2009). https://doi.org/10.1007/s11105-008-0077-9

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