Genotyping of bovine k-casein (k-CNA, k-CNB, k-CNC, k-CNE) following DNA sequence amplification and direct sequencing of k-CNE PCR product |
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Authors: | S SCHLIEBEN G ERHARDT B SENFT |
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Institution: | Institut für Tierzucht und Haustiergenetik, Justus-Liebig-Universit?t, Giessen, Germany. |
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Abstract: | 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. |
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Keywords: | bovine k-casein genetic polymorphism polymerase chain reaction direct sequencing genotyping |
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