Analysis of the Genotype Distribution in Cattle Breees Using a Double Mismatched Primer Set that Discriminates the MC1R Dominant Black Allele

摘要

With a double mismatch primer set designed for amplifying the modified DNA sequence fragments, bovine melanocortin-1 receptor (MC1R) gene encoded in Extension locus which plays a critical role in coat color development was analyzed using polymerase chain reaction mediated restriction fragment length polymorphism (PCR-RFLP). Amplified PCR fragments were successfully discriminated with combining the Mspl- and Alul-RFLP into three major alleles (E~D, E~+, and e), directly related to bovine coat color phenotypes. The genotyping results showed that Jeju black cattle contained three MC1R alleles, but yellowish-red colored Hanwoo and bridle colored Korean Brindle cattle did not contained the dominant black allele if. However, two dominant black-colored cattle breeds, Holstein and Angus, contained the E~D allele over 96% in frequency. HanwooxHolstein FI and HanwooxAngus Fi crossbred calves showed E~D/e MC1R genotypes, and uniformly black coat color, the results suggested that this MC1R genotyping method beuseful in allele discrimination for bovine MC1R gene which used for breed classification and characterization, as one of the important genetic markers, using combination of Mspl- and .AluI-RFLP for modified PCR product amplified with a newly designed double mismatch primer set.