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. Assignment of the SIAT4A gene to bovine chromosome 14 by linkage mapping of an associated microsatellite. Animal Genetics 35:146-147. Interpretive Summary: A new DNA marker for the Sialyltransferase 4A (SIAT4A) gene was developed and mapped. The marker was developed from the CHORI-240 bacterial artificial chromosome library. Cattle from an Angus-based commercial seedstock line, and two USDA-MARC reference families were genotyped. The USDA-MARC reference family genotypes were used to map the marker onto cattle chromosome 14.
Technical Abstract: CHORI-240 bovine bacterial artificial chromosome library high density filters were probed with gene-specific overgo primers for Sialyltransferase 4A (SIAT4A). All positive clones were confirmed by polymerase chain reaction (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, and sequences were queried through Blastn searches against GenBank. Oligonucleotide primers for this sequence were designed, and the microsatellite marker was amplified via PCR then analysed using a denaturing polyacrylamide gel electrophoresis system. Polymorphism was examined using 213 parents from the M1 commercial line of Beefbooster Inc. (Calgary, Canada) and two USDA-MARC cattle reference families. 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 locus UASM014 was mapped onto bovine chromosome 14 at approximately 5.7 cM from marker BM1508. On the basis of its colocation with UASM014 in the BAC clone the SIAT4A locus can also be mapped to BTA 14.