Schlieben S, Erhardt G, Senft B
Institut für Tierzucht und Haustiergenetik, Justus-Liebig-Universität, Giessen, Germany.
Anim Genet. 1991;22(4):333-42. doi: 10.1111/j.1365-2052.1991.tb00687.x.
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.
从已知κ-酪蛋白（κ-CN）基因型的奶牛血液和精液中分离出的基因组DNA，使用多达14种限制性内切酶进行Southern印迹杂交和聚合酶链反应（PCR）。使用Hin dIII和Hin fI鉴定出κ-酪蛋白基因型AA、AB和BB，而κ-CNC和κ-CNE基因型被误鉴定。PCR产物（κ-CN EE）的直接测序显示鸟嘌呤（κ-CNA,B）被腺嘌呤（κ-CNE）取代，这产生了一个HaeIII限制性位点。因此，采用PCR后进行Hin dIII或HinfI和Hae III酶切，能够在DNA水平直接区分κ-酪蛋白A、B和E。
