Biochemical characterization of bovine alpha s1-casein F and genotyping with sequence-specific primers.

Bovine alpha s1-casein F (alpha s1-CN F) was found in a genetic resource of Deutsches Schwarzbuntes Niederungsrind cows at a frequency of 0.009. Biochemical characterization of this new variant was obtained by automated sequencing of reversed-phase HPLC-separated tryptic peptides of alpha s1-CN F and alpha s1-CN B. alpha s1-CN F was found to be a subtype of alpha s1-CN B with a single amino acid substitution (SerP/Leu) in position 66. DNA sequencing revealed a C/T transition in position 8418 of the gene. Sequence-specific primers were designed to perform an allele-specific polymerase chain reaction for detection of alpha s1 CnF. Typing of artificial insemination sperm samples included in the genetic resource sperm pool identified one sire heterozygous for alpha s1 CnF.