The polymerase chain reaction (PCR) was used to amplify a 432 bp region from the k- casein gene of Holstein dairy cattle which contains the nucleotide substitutions diagnostic of the 2 major protein variants of K-casein. Digestion of the PCR product with Hinfl (A variant specific) or TaqI (B variant specific) allowed direct determination of the genotype of the animal (homozygous or heterozygous). The same samples were also analyzed using single strand conformation polymorphism analysis (SSCP) PCR. Full length (432 bp) amplicons and smaller fragments were analyzed in order to increase the probability of detecting DNA polymorphisms in addition to the 2 characterized above. Analysis of cows (n=123) and bull calves (n=102) revealed no difference in frequency of the A allele (.81 and .78, respectively), whereas selected sires had a frequency of .87 for this allele. Analysis of 60 animals (to date) using SSCP has revealed no additional changes in genetic structure of exon IV of K-casein but did detect the DNA polymorphisms associated with the allelotypes. The major advantage over RFLP analysis is that SSCP-PCR has the potential to detect additional genetic variants in the population
Proceedings of the World Congress on Genetics Applied to Livestock Production, Volume 21. Gene mapping; polymorphisms; disease genetic markers; marker assisted selection; gene expression; transgenes; non-convention, , 140–143, 1994
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