Caseins are a family of milk proteins that exist in several molecular forms (alphas1, alphas2, beta and kappa) with variant alleles of each. Kappa-caseins (κ-CN) exist as A or B variants and are of particular importance to the quality of the milk. We report a rapid non-radioactive method for differentiating bovine k-CN alleles. Unlike direct milk protein analysis, this method can be utilized for genotyping sperm, or animals of either sex at an early age. A 350bp fragment of the bovine κ-CN gene was amplified by the polymerase chain reaction (PCR) technique. The amplified product digested with Hinfl endonuclease generated restriction fragment length polymorphisms (RFLP's) that allowed precise identification of κ-CN AA, AB or BB genotypes. Hybridization with allele specific oligo-probes confirmed the RFLP identification of genotypes. The accurate and early identification of milk protein genotypes can have a direct impact on dairy cattle breeding strategies.
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Eigel, W.N., Butler, J.E., Ernstrom, C.A., Farrel, Jr., H.M., Harwalkar, V.R., Jenness, R. and Whitney, R. Mcl. 1984. Nomenclature of proteins of cow's milk: Fifth Revision. J. Dairy Sci. 67: 1599–1631.
Grosclaude, F. 1988. Le polymorphisme génétique des principales lactoproteines bovines. INRA Prod. Anim. 1: 5–17.
Russo, V. and Mariani, P. 1978. Polimorfismo delle proteine del latte e relazioni tra varianti genetiche e caratteristiche di interesse zootecnico tecnologico e caseario. Revista di Zootecnia e Veterinaria 5: 1–31.
Schaar, J., Hansson, B. and Pettersson, H.E. 1985. Effects of genetic variants of κ-casein and β-lactoglobulin on cheesemaking. J. Dairy Res. 52: 429–437.
Wake, R.G. and Baldwin, R.L. 1961. Analysis of casein fractions by zone electrophoresis in concentrated urea. Biochim Biophys. Acta 47: 225–239.
Aschaffenburg, R. 1961. Inherited casein variants in cow's milk. Nature 192: 431–432.
Mackinlay, A.G. and Wake, R. 1971. κ-casein and its attack by rennin (chymosin). In: Milk Proteins, Chemistry and Molecular Biology. H.A. Mckenzie (Ed.). Academic Press, NY.
Groves, M.L. 1969. Some minor components of casein and other phosphoproteins in milk: A review. J. Dairy Sci. 52: 1155–1165.
Neelin, J.M. 1964. Variants of κ-casein revealed by improved starch gel electrophoresis. J. Dairy Sci. 47: 506–509.
Schmidt, D.G. 1964. Starch gel electrophoresis of κ-casein. Biochim. Biophys. Acta 90: 411–414.
Gorodetskii, S.I., Kershulyte, D.R. and Korobko, V.G. 1983. Nucleotide sequence of cDNA of kappa-casein macropeptide of bos taurus. Bioorganicheskaya Khimia, USSR 9: 1693–1695.
Kang, Y. and Richardson, T. 1988. Molecular cloning and expression of bovine κ-casein in Escherichia coli . J Dairy Sci. 71: 29–40.
Erlich, H.A., Gelfand, D.H. and Saiki, R.K. 1988. Specific DNA amplification. Nature 331: 461–462.
Alexander, L.J., Stewart, A.F., Mackinlay, A.G., Kapelinskaya, T.V., Tkach, T.M. and Gorodetsky, S.I. 1988. Isolation and characterization of bovine κ-casein gene. J. Biochem. 178: 395–401.
Aschaffenburg, R. 1968. Genetic variation of milk proteins. J. Dairy Res. 35: 447–460.
Madisen, L., Hoar, D.I., Holroyd, C.D., Crisp, M. and Hodes, M.E. 1987. DNA banking: the effects of storage of blood and isolated DNA on the integrity of DNA. Am. J. Med. Genetics 27: 379–390.
Higuchi, R., 1989. Rapid, efficient DNA extraction for PCR from cells or blood. Perkin Elmer/Cetus Newsletter. Amplifications 2: 1–3.
Gebeyehu, G., Rao, P.Y., SooChan, P., Simms, D.A. and Klevan, L. 1987. Novel biotinylated nucleotide—analogs for labeling and colorimetric detection of DNA. Nucl. Acids Res. 15: 4513–4534.
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Medrano, J., Cordova, E. Genotyping of Bovine Kappa-Casein Loci Following DNA Sequence Amplification. Nat Biotechnol 8, 144–146 (1990). https://doi.org/10.1038/nbt0290-144
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