Submitted to: Animal Genetics
Publication Type: Peer reviewed journal
Publication Acceptance Date: 12/5/2003
Publication Date: 4/1/2004
Citation: Murdoch, B., Fu, A., Meng, Y., Li, C., Hansen, C., Snelling, W.M., Moore, S.S. 2004. Identification of three microsatellite loci on bovine chromosome 19. Animal Genetics 35:145-146. Interpretive Summary: Three new DNA markers were developed and mapped on cattle chromosome 19. The markers were developed from the CHORI-240 bovine bacterial artificial chromosome library using gene-specific probes for thyroid hormone receptor alpha (THRA), acetyl-coenzyme A carboxylase alpha (ACACA), and insulin-like growth factor-binding protein 4 (IGFBP4). Cattle from an Angus-based commercial seedstock line, and two USDA-MARC reference families were genotyped for the new markers associated with each gene. The USDA-MARC reference family genotypes were used to map the markers onto cattle chromosome 19. Placements of the markers on chromosome 19 agree with placement expected from the human-bovine comparative map.
Technical Abstract: CHORI-240 bovine bacterial artificial chromosome (BAC) library high density filters were probed with gene-specific overgo primers for thyroid hormone receptor alpha (THRA), acetyl-coenzyme A carboxylase alpha (ACACA), and insulin-like growth factor-binding protein 4 (IGFBP4). All positive clones were confirmed by PCR with different gene-specific primers. Subsequently the clones were digested with Sau3 AI and subcloned into the E. coli cloning vector pUC18. Dinucleotide repeats were identified by screening colony hybridization with a radioactive labeled polynucleotide, alternating poly (dA-dC) and poly (dT-dG). The positive subclones were sequenced with the use of both the forward and reverse universal primers. The sequences were queried through NCBI GenBank Blastn searches. Oligonucleotide primers for these sequences were designed, and the microsatellites were amplified via PCR. Polymorphisms at each locus were examined using 213 parents from the M1 commercial line of Beefbooster, Inc. (Calgary, Canada) and two USDA-MARC cattle reference families. The genotypes of two USDA-MARC cattle reference families were used for the determination of genetic map locations for the microsatellite loci. Chromosomal assignments of microsatellite loci were based on sex-averaged two-point LOD scores >3.0 using CRI-MAP version 2.4. Microsatellite loci UASM010, UASM011 and UASM013 were all mapped onto bovine chromosome 19, as expected based on the prediction from the human-bovine comparative map, approximately 0.5 cM from BMS650, 2.5 cM from RM222 and 2.7 cM from CSSM065, respectively.