ARS | Publication request: CONSTRUCTION OF A COTTON BAC LIBRARY AND ITS APPLICATION TO GENE ISOLATION

Submitted to: Plant and Animal Genome
Publication Type: Abstract Only
Publication Acceptance Date: January 6, 2000
Publication Date: January 6, 2000
Citation: Yu, J., Kohel, R.J., Zhang, H., Dong, J., Decanini, L. 2000. Construction of a cotton BAC library and its application to gene isolation [abstract]. Plant and Animal Genome VIII Conference. Paper No. P146.

Technical Abstract: Cotton (Gossypium spp.) is a leading natural fiber crop. This crop is also an excellent source of the oil and protein that are stored in its seed. As a major crop species in the world, cotton has a potentially broad genetic base, reflected in the collections of Gossypium species. Cotton breeders would have therefore an opportunity to exploit and manipulate many thousand genes in the Gossypium reservoir. To isolate genes in cotton via positional cloning, high-quality libraries are needed that are comprised of bacterial artificial chromosome (BAC) clones containing large cotton DNA inserts. We are currently in the process of constructing such a BAC library from a Gl2e-BC6 nearly isogenic line (NIL) of G. hirsutum acc. TM-1. An average size of the DNA inserts is about 150 kb, with a cloning site of BamHI, EcoRI, and HindIII (2 X haploid genome equivalents each enzyme), respectively. DNA markers flanking Gl2e, the glandless gene, have been identified via linkage analysis of an F2 population derived from the cross between the TM-1 NILs (with and without Gl2e). They are used to screen the BAC library for the isolation of the Gl2e gene, and additional DNA markers are developed from the positive BAC clones. Other genes such as resistance to the diseases tan nemotodes in cotton are also taking advantage of this BAC library for positional cloning. Details in the construction, characterization, and general applications of the BAC library will be provided in the report.