|Ponce De Leon, F|
Submitted to: Cytogenetics and Genome Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/19/2002
Publication Date: 6/28/2002
Citation: Reed, K.M., Ihara, N., Mariani, P., Mendoza, K.M., Jensen, L.E., Bellavia, R., Ponce De Leon, F.A., Bennett, G.L., Sugimoto, Y., Beattie, C.W. 2002. High-resolution genetic map of bovine chromosome 29 through focused marker development. Cytogenetics and Genome Research. 96:210-216. Interpretive Summary: Having many markers is very useful when looking for the location of genetic differences on chromosomes. Researchers can choose markers with good locations, clear marker genotypes, and identifiable forms in the populations being studied. A method for increasing the number of markers on a specific chromosome was used to nearly triple the markers on cattle chromosome 29. The method had about an 85 percent success rate with 15 percent of newly developed markers mapped to chromosomes other than chromosome 29. Results show that the method can be successful. Results have also yielded a useful resource for refining the estimated location of genetic variation on cattle chromosome 29.
Technical Abstract: Chromosome-specific libraries aid in the development of genetic maps and focus marker development in areas of the genome with identified QTL. A small-insert BTA29 library constructed by microdissection of a 1:29 Rb-fusion cell line, was screened for dinucleotide repeats (CA) 15 and/or (GA) 15. Approximately 1200 primary clones were recovered and secondarily screened. DNA sequences were determined for 458 positive clones. From these, a total of 90 primer pairs were designed and 82 of these successfully amplified bovine genomic DNA by PCR. In addition to these 82 loci, primer pairs were developed for nine putative genes identified from the sequenced clones by BLAST searches of Genbank. A somatic cell panel was used to test for synteny of the new loci with two previously mapped BTA29 markers located on the MARC bovine linkage map (BMC2228 and BMC3224). Results of these tests show that 85% of the loci are syntenic with the previously mapped loci. Two loci were linked to markers on BTA1 and nine loci did not map to either BTA29 or BTA1 (four of these constitute a separate linkage group). A total of 75 of the 82 microsatellite loci were also integrated into the MARC bovine linkage map. Linkage analysis placed 69 ms markers on BTA29, five on BTAX and one on BTA1. Combined results of the somatic cell and linkage analyses place 79 new markers (ms and gene-related) on BTA29, six loci on BTAX and two loci on BTA1. The results of this effort significantly increase the marker density on BTA29.