Genotyping of bovine kappa-casein (kappa-CNA, kappa-CNB, kappa-CNC, kappa-CNE) following DNA sequence amplification and direct sequencing of kappa-CNE PCR product.
Genomic DNA isolated from blood and semen of dairy cattle with known kappa-casein (kappa-CN) genotypes was subjected to Southern blot hybridization and polymerase chain reaction (PCR) using up to 14 restriction endonucleases. kappa-casein genotypes AA, AB and BB were identified using Hin dIII and Hin fI while genotypes with kappa-CNC and kappa-CNE were misidentified. Direct sequencing of the PCR product (kappa-CN EE) showed a substitution of guanine (kappa-CNA,B) by adenine (kappa-CNE) which creates a HaeIII restriction site. Therefore using PCR followed by Hin dIII or HinfI and Hae III digest allows discrimination between kappa-casein A, B and E directly at the DNA level.
Institut für Tierzucht und Haustiergenetik, Justus-Liebig-Universität, Giessen, Germany.
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Schlieben
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Erhardt
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Senft
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002364:Caseins / Q000235:genetics*; D002417:Cattle / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D005091:Exons / Q000235:genetics; D005838:Genotype; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
D002364:Caseins; D009838:Oligodeoxyribonucleotides; D004262:DNA Restriction Enzymes