Two large-insert soybean genomic libraries constructed in a binary vector: applications in chromosome walking and genome wide physical mapping Academic Article uri icon

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 BamHI or HindIII. 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.

published proceedings

  • Theoretical and Applied Genetics

altmetric score

  • 3

author list (cited authors)

  • Meksem, K., Zobrist, K., Ruben, E., Hyten, D., Quanzhou, T., Zhang, H., & Lightfoot, D. A.

citation count

  • 63

complete list of authors

  • Meksem, K||Zobrist, K||Ruben, E||Hyten, D||Quanzhou, T||Zhang, H-B||Lightfoot, DA

publication date

  • January 1, 2000 11:11 AM