CHROMOSOME SPECIFIC LIBRARIES AND THEIR USE IN GENOME MAPPING

Authors: Lunney, Joan; Grimm, David; Holley Shanks, Rhonda; GOLDMAN, T - USDA; PONCE DE LEON, F - UNIV MASSACHUSETTS; AMBADY, S - UNIV MASSACHUSETTS; ROBL, J - UNIV MASSACHUSETTS; MENDIOLA, J - UNIV MINNESOTA; BUOEN, L - UNIV MINNESOTA; LOUIS, C - UNIV MINNESOTA

Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: 5/10/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Swine chromosome specific libraries have been prepared by our laboratories using both flow cytometric and microdissection techniques. Such libraries have been successfully used in swine and other species to expand the number of genomic markers available, particularly for chromosomes, or areas of chromosomes, that are linked to important traits. Our target has been to produce more swine chromosome 6 microsatellite markers so that we can identify unique loci near the porcine stress syndrome defect that determine favorable swine carcass traits. For flow cytometry, chromosomes were prepared from activated lymphoblasts and sorted based on their staining with Hoechst 33258 and chromomycin A3. Libraries were produced from size selected DNA fragments amplified from fewer than 1,000 sorted chromosomes. Chromosomes have also been isolated by microdissection techniques and then PCR amplified using specifically designed primers. Purity of these populations was verified by fluorescent in situ hybridization (FISH) analyses of painting probes. The swine libraries were screened with dinucleotide repeat probes and, after sequencing, PCR primers designed for each marker, with the result that many chromosome 6 microsatellite specific markers have been identified and mapped. The development of chromosome enriched libraries has enabled many new markers to be isolated that can be used to identify the actual genes that influence production traits.