|GRIMM DAVID R|
Submitted to: BARC Poster Day
Publication Type: Abstract Only
Publication Acceptance Date: 2/22/1994
Publication Date: N/A
Technical Abstract: A swine chromosome 6 specific library was generated using size fractionated DNA isolated from chromosomes sorted by flow cytometry. Pig chromosome 6 is important because the genes that encode positive carcass traits, such as lean meat content, loin eye muscle area, and decreased back fat depth, have been mapped to this chromosome. Our chromosome 6 library will be used to develop new microsatellite markers to separate these positive carcass traits from the negative trait, porcine stress syndrome, which causes sudden death in pigs and is associated with soft exudative meat. Chromosome isolation procedures have been established in our laboratory to reproducibly prepare high quality chromosomes and to sort individual chromosomes after staining with Hoechst 33258 and chromomycin A using a Coulter Epics flow cytometer. DNA from sorted chromosome 6 was digested with Sau3A and ligated into pBluescript SK+. After PCR amplification using the T7 and T3 primers, DNA fragments of 300-600 base pairs were size fractionated on agarose gels, religated into pBluescript SK+ and used to transform DH5-alpha E. coli cells. This chromosome 6 library was then searched for the presence of dinucleotide microsatellite markers using a biotinylated (GT)15 oligonucleotide for the detection of (CA)n microsatellite repeats. After sequencing, these new genetic markers on chromosome 6 will enable us to begin to develop a more detailed map of the region where these genes are found. Herds of pigs, that have been characterized for their carcass traits and inheritance of the porcine stress syndrome allele, will be analyzed with these markers for alleles that will identify genetically superior animals for future breeding stock.