PAG-VI: ESTABLISHMENT OF A BAC LIBRARY RESOURCE CENTER FOR AGRICULTURAL GENOMICS

PAG-VI  Plant & Animal Genome VI Conference

Town & Country Hotel, San Diego, CA, January 18-22, 1998.

W88,W124

ESTABLISHMENT OF A BAC LIBRARY RESOURCE CENTER FOR AGRICULTURAL GENOMICS

DAVID A FRISCH1, Dilara Begum1, Muhammad Budiman1, Sangdun Choi1, Jeff Tomkins1, Robbie Waugh2, Yeisoo Yu1, Heng Zhu1, Maciek Sasinowski1, Ralph Dean1, Rod Wing1

  1. Clemson University Genome Center, Clemson University, Clemson, SC 29634
  2. Scottish Crop Research Institure, Mylnefield, Invergowrie, Dundee DD25DA

The CU Genome Center (http://genome.clemson.edu) has established a large insert BAC library service facility for agricultural genomics. The functions of the facility are:

  1. Construction and maintenance of high quality BAC libraries.
  2. Production and distribution of high density hybridization filters and DNA pools for library screening and distribution of BAC libraries and clones.
  3. Positional cloning of agriculturally important genes.
  4. Physical mapping of whole chromosomes and targeted genomic regions with BAC clones.
  5. Genome sequencing of important BAC contigs and genomes.
  6. Application development.
  7. Undergraduate, graduate and postdoctoral training in genomics.
Since 1993, our laboratory has produced and distributed a large number of BAC libraries. We have produced representative libraries for crop plants such as tomato, sugarcane, sorghum, rice, and barley. A large insert BAC library has also been produced for Magnaportha grisea, the fungal agent that causes rice blast. The most recent libraries produced are from barley (5000 Mb genome, 2X coverage), tomato (950 Mb genome, 3X coverage), sugarcane (3000 Mb genome, 3X coverage) and citrus (380 Mb, 5X coverage). Work is continuing to expand the barley library to 6X and the tomato library to 15X. The genome center has developed procedures to efficiently produce and screen BAC libraries using the multitasking Q-Bot. We routinely produce 22.5 cm2 filters containing with forty eight 384-well plates spotted in duplicate (18,432 unique clones). Hybridization of these filters allows for the easy identification of individual BAC clones containing the sequence of interest. We are presently using the system to produce filters for the TAMU and IGF A. thaliana BAC libraries being used for the A. thaliana genome sequencing project and distributed by ABRC. Details will be presented on the construction of BAC libraries and results obtained from library screenings. The CU Genome Center is funded in part by Clemson University, the Robert and Lois Coker Endowment the National Science Foundation and USDA-NRICGP. The CU Genome Center would like to thank T. Altmann (Arabidopsis), S. Mackenzie (common bean), R. Shoemaker (soybean), P. Ronald (Arabidopsis, rice), N. Young (soybean) for donating their BAC libraries for distribution through the CU Genome Center.

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