Two Large-Insert Soybean Genomic Libraries Constructed in a Binary Vector: Applications in Chromosome Walking and Genome Wide Physical Mapping

Authors

K. Meksem, Southern Illinois UniversityFollow
K. Zobrist, Southern Illinois University
E. Ruben, Southern Illinois University
D. L. Hyten, Southern Illinois UniversityFollow
T. Quanzhou, Texas A&M University
H-B. Zhang, Texas A&M UniversityFollow
D. A. Lightfoot, Southern Illinois UniversityFollow

ORCID IDs

D. L. Hyten

Document Type

Article

Date of this Version

10-2000

Citation

Theoretical and Applied Genetics 101:5–6 (October 2000), pp. 747–755

doi: 10.1007/s001220051540

Comments

Copyright © 2000 Springer-Verlag. Used by permission.

Abstract

Large DNA insert libraries in binary T-DNA vectors can assist in the isolation of the gene(s) under-lying 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.