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

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 art...

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Bibliographic Details
Published in:Plant molecular biology reporter 2009-06, Vol.27 (2), p.229-235
Main Authors: Zhu, S, Saski, C. A, Boerma, H. R, Tomkins, J. P, All, J. N, Parrott, W. A
Format: Article
Language:English
Subjects:
Artificial chromosomes
Bacteria
Bacterial artificial chromosomes
Bioinformatics
Biomedical and Life Sciences
Cloning
Deoxyribonucleic acid
DNA
DNA libraries
Gene mapping
Gene sequencing
genetic markers
Genomes
Genotypes
Glycine max
haploidy
insect pests
Insects
Life Sciences
linkage (genetics)
Metabolomics
molecular cloning
new methods
Nucleotide sequence
pest resistance
Plant Breeding/Biotechnology
plant pests
Plant Sciences
plant-insect relations
Proteomics
Quantitative trait loci
repetitive sequences
Soybeans
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Summary: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.
ISSN:0735-9640
1572-9818
DOI:10.1007/s11105-008-0077-9