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.
Copyright policy )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.
- Justus Liebig University Giessen Germany
Base Sequence, Genotype, Molecular Sequence Data, Caseins, DNA Restriction Enzymes, Exons, Polymerase Chain Reaction, Introns, Blotting, Southern, Oligodeoxyribonucleotides, Animals, Cattle, Polymorphism, Restriction Fragment Length
Base Sequence, Genotype, Molecular Sequence Data, Caseins, DNA Restriction Enzymes, Exons, Polymerase Chain Reaction, Introns, Blotting, Southern, Oligodeoxyribonucleotides, Animals, Cattle, Polymorphism, Restriction Fragment Length
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