Sulimova G E, Shaĭkhaev G O, Berberov E M, Markarian A Iu, Kandalova L G
Genetika. 1991 Dec;27(12):2053-62.
Polymerase chain reaction (PCR) was used for detecting kappa-casein (kappa-casein) genotype in the synthetic breed (Jersey x Black and White x Holstein-Friesian) dairy cattle. The amplified 228 bp fragment includes a region, where relevant mutations lead to both the appearance of different kappa-casein alleles associated with amino acid substitutions and the appearance of new TaqI and HindIII restriction sites in ae-casein B gene. The specificity of the kappa-casein gene fragment amplification was supported by restriction analysis and Southern blot hybridization. Digestion of amplified fragment with endonucleases PstI, HindIII and/or TaqI allows detection of AA-, AB- and BB genotypes of kappa-casein. A total of 32 animals with known (18 samples) and unknown (14 samples) kappa-casein phenotypes were tested using PCR and blot hybridization. In all known cases the detected genotype confirmed the phenotype. Frequencies of the B allele and of the AB genotype in the breeding population are rather high (53.1 +/- 8.8 and 43.7%, respectively). The possibility of effective use of the PCR analysis for genotyping kappa-casein locus in bulls and their offspring has been shown. The advantages of the PCR method in large breeding programs and linkage analysis have been discussed.
采用聚合酶链反应（PCR）检测合成品种（泽西牛×黑白花牛×荷斯坦-弗里生牛）奶牛的κ-酪蛋白（kappa-casein）基因型。扩增得到的228 bp片段包含一个区域，相关突变导致与氨基酸替换相关的不同κ-酪蛋白等位基因出现，以及在αs1-酪蛋白B基因中出现新的TaqI和HindIII限制性酶切位点。κ-酪蛋白基因片段扩增的特异性通过限制性酶切分析和Southern印迹杂交得到证实。用限制性内切酶PstI、HindIII和/或TaqI消化扩增片段可检测κ-酪蛋白的AA、AB和BB基因型。使用PCR和印迹杂交对总共32头已知（18个样本）和未知（14个样本）κ-酪蛋白表型的动物进行了检测。在所有已知病例中，检测到的基因型与表型相符。育种群体中B等位基因和AB基因型的频率相当高（分别为53.1±8.8和43.7%）。已经证明了有效利用PCR分析对公牛及其后代的κ-酪蛋白基因座进行基因分型的可能性。讨论了PCR方法在大型育种计划和连锁分析中的优势。
