Plant & Animal Genome IX Conference

Poster: Large Insert Libraries, Gene Isolation, Etc.
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BACTERIAL ARTIFICIAL CHROMOSOME (BAC) LIBRARIES CONSTRUCTED FROM THE GENETIC STANDARD OF UPLAND COTTONS

¹ USDA-ARS, Crop Germplasm Research Unit, 2765 F&B Road, College Station, TX 77845, USA

² Texas A&M University, Department of Soil and Crop Sciences, College Station, TX 77843, USA

Two BAC libraries and one plant transformation-competent BIBAC library were developed from the Gossypium hirsutum acc. TM-1 for the development of an integrative cotton physical and genetic map and other genomic applications. TM-1 is the most desirable choice for construction of the physical map of Upland cotton because it is highly homozygous, maintained by single seed decent in the 52^nd selfing generation, which is required for genome-wide physical mapping by fingerprint analysis of large insert clones. Furthermore, this line was extract from a widely adapted and productive cultivar, and it is widely used as the genetic standard of Upland cottons (G. hirsutum L.). Many genetic and cytogenetic stocks are produced in the TM-1 background and TM-1 has been used as one of the parents of recombinant inbred line populations for cotton genome mapping. A high-density DNA marker map has been developed from the population. The two BAC libraries were constructed in the HindIII and EcoRI sites of pECBAC1, respectively. The small size of pECBAC1 (7.4 Kb) will facilitate the use of the BAC clones for genome analysis and local genome sequencing. A BIBAC library was constructed in the BamHI site of the plant transformation-competent binary vector pCLD04541. The Agrobacterium-mediated plant transformability of large-insert pCLD04541 clones streamlines positional cloning of the genes, QTLs, gene engineering, and molecular breeding by transformation. These three TM-1 BAC and BIBAC libraries contain 172,800 BAC clones (57,600 clones for each library), which should be sufficient for development of a genome-wide BAC/BIBAC-based physical map of the Upland cotton genome. These libraries are complementary to each other since they were constructed in different vector systems and with three complementary enzymes in restriction site sequences, and they are equivalent to a physically sheared shotgun library in the distribution of clones in the genome.