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The identification of the kappa-casein genotype in Holstein dairy cattle using the polymerase chain reaction

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Summary

The polymerase chain reaction (PCR) was used to amplify a 99-bp region from the kappa-casein gene of Holstein dairy cattle which contains nucleotide substitutions that are diagnostic of the two major protein variants of kappa-casein. Identity of the amplified product was confirmed by direct sequencing. Digestion of the PCR product with MboII (A-variant specific) or TaqI (B-variant specific) allowed direct determination of the genotype of the animal (homozygous or heterozygous). A total of 58 lactating cows with known kappa-casein phenotype were tested using PCR. In all cases, the measured genotype confirmed the phenotype. We have also tested the genotype of 42 sires that were top ranked for milk yield by the CIAQ (Centre d'insemination artificielle du Quebec). The B-allele of kappa-casein which occurred at a frequency of 0.13 among the proven bulls is associated with superior milk for industrial applications. Identification of the kappa-casein genotype by PCR in bulls and calves would provide a means for rapidly changing the frequency of the B-allele in the breeding population by selection.

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References

  • Alexander LJ, Stewart AF, MacKinlay AG, Kapelinskaya TV, Tkatch TM, Gorodetsky SI (1988) Isolation and characterization of the bovine kappa-casein gene. Eur J. Biochem 178:395–401

    Google Scholar 

  • Feldman GL, Williamson R, Beaudet AL, O'Brian WE (1988) Prenatal diagnosis of cystic fibrosis by DNA amplification for detection of KM-19 polymorphism. Lancet ii: 102

    Google Scholar 

  • Gyllensten UB, Erlich HA (1988) Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus. Proc Natl Acad Sci 80:7652–7656

    Google Scholar 

  • Kang YC, Richardson T (1988) Molecular cloning and expression of bovine kappa-casein in Escherichia coli. J. Dairy Sci 71:29–40

    Google Scholar 

  • Kogan SC, Doherty M, Gitsshier J (1987) An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences: application to hemophilia A. N Engl J Med 317:985–990

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y

    Google Scholar 

  • Marziali AS, Ng-Kwai-Hang KF (1986) Relationships between milk protein polymorphisms and cheese yielding capacity. J Dairy Sci 69:1193–1201

    Google Scholar 

  • McLean DM (1987) Influence of milk protein variants on milk composition, yield and cheesemaking properties. Anim Genet 18 [Suppl 1]

  • McLean DM, Graham ERB, Ponzoni RW, McKenzie HA (1984) Effects of milk protein genetic variants on milk yield and composition. J Dairy Res 51:531–546

    Google Scholar 

  • Medrano JF, Aguilar-Cordova E (1990) Genotyping of bovine kappa-casein loci following DNA sequence amplification. Biotechnology 8:144–146

    Google Scholar 

  • Mercier JC, Brignon G, Rimbadeau-Dumas B (1973) Structure primaire de la casein kappa-B bovine. Sequence complete. Eur J Biochem 23:41–45

    Google Scholar 

  • Morini D, Losi G, Castagnetti GB (1979). Properties of ripened cheese in cheesemaking experiments with milk characterized by kappa-casein variants A and B. Sci Tec Lattiero-Casearia 30:1–20

    Google Scholar 

  • Ng-Kwai-Hang KF, Kroeker EM (1984) Rapid separation and quantification of major caseins and whey proteins of bovine milk by polyacrylamide gel electrophoresis. J Dairy Sci 67:3052–3056

    Google Scholar 

  • Ng-Kwai-Hang KF, Hayes JF, Moxley JE, Monardes HG (1984) Associations of genetic variants of casein and milk serum proteins with milk, fat and protein production in dairy cattle. J Dairy Sci 67:835–840

    Google Scholar 

  • Rando A, DiGregorio P, Masina P (1988) Identification of bovine K-casein genotypes at the DNA level. Anim Genet 19:51–54

    Google Scholar 

  • Saiki RT, Bugawan TL, Horn GT, Mullis KB, Erlich HA (1986) Analysis of enzymatically amplified beta-globin and HLADQ-alpha DNA with allele-specific oligonucleotide probes. Nature 324:163–166

    Google Scholar 

  • Tsang TC, Bentley DR, Mibashan RS, Giannelli F (1988). A factor IX mutation, verified by direct genomic sequencing, causes hemophilia B by a novel mechanism. EMBO J 7:3009–3015

    Google Scholar 

  • Wrischnik LA, Higuchi RG, Stoneking M, Erlich HA, Arnheim N, Wilson AC (1987) Length mutations in human mitochondrial DNA: direct sequencing of enzymatically amplified DNA. Nucleic Acids Res 15:529–542

    Google Scholar 

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Communicated by K. Sitmann

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Zadworny, D., Kuhnlein, U. The identification of the kappa-casein genotype in Holstein dairy cattle using the polymerase chain reaction. Theoret. Appl. Genetics 80, 631–634 (1990). https://doi.org/10.1007/BF00224222

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  • DOI: https://doi.org/10.1007/BF00224222

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