Sulimova G E, Badagueva Iu N, Udina I G
Genetika. 1996 Nov;32(11):1576-82.
Polymorphism of the 5'-untranslated region and exon 4 of kappa-casein (kappa-casein) gene was studied in Yakutian and Black Pied cattle, yak, European bison, and buffalo by means of a polymerase chain reaction and subsequent restriction fragment length polymorphism analysis (PCR-RFLP). In the species studied, restriction polymorphism by the endonucleases AluI and Bg/II in the 5'-untranslated region of the gene is absent. Four restriction endonucleases testing nucleotide substitutions in 136 codon (TaqI), 148 codon (HinfI and HindIII), and 167 and 168 codons (PstI) were used to study polymorphism of exon 4. The use of several restriction endonucleases allowed three alleles of kappa-casein (kappa-CnA, kappa-CnB, kappa-CnF) to be typed and new allele variants in yak, European bison, and buffalo to be revealed. Nucleotide sequences of the fragments of exon 4 studied were determined for two new alleles of the gene: kappa-CnG in yak and European bison and kappa-CnH in buffalo. Nucleotide substitutions determining new alleles were localized. In kappa-CnG, 148 and 168 codons coincide with the corresponding codons of kappa-CnB, and 136 and 167 codons correspond to kappa-CnA. Stop codons of kappa-CnG in yak are different from stop codons of other alleles of the gene: TGA, instead of TAA. The nucleotide sequence of exon 4 of kappa-CnH differs from bovine kappa-CnA by 15 nucleotide substitutions, causing 10 amino acid changes in the protein sequence, which coincide with the corresponding known amino acid sequence of kappa-casein in buffalo. Interbreed and interspecies differences in the profile of allele frequencies of the species studied were revealed. Aspects connected with evolution of the alleles of kappa-casein are discussed.
采用聚合酶链反应及随后的限制性片段长度多态性分析(PCR-RFLP),对雅库特牛、黑花斑牛、牦牛、欧洲野牛和水牛的κ-酪蛋白(kappa-casein)基因5'-非翻译区和外显子4的多态性进行了研究。在所研究的物种中,该基因5'-非翻译区不存在AluI和Bg/II内切酶的限制性多态性。使用4种限制性内切酶检测第136密码子(TaqI)、第148密码子(HinfI和HindIII)以及第167和168密码子(PstI)处的核苷酸替换,以研究外显子4的多态性。使用多种限制性内切酶使得能够对κ-酪蛋白的3个等位基因(κ-CnA、κ-CnB、κ-CnF)进行分型,并揭示了牦牛、欧洲野牛和水牛中的新等位基因变体。测定了该基因外显子4片段的核苷酸序列,确定了两个新等位基因:牦牛和欧洲野牛中的κ-CnG以及水牛中的κ-CnH。确定了决定新等位基因的核苷酸替换位置。在κ-CnG中,第148和168密码子与κ-CnB的相应密码子一致,第136和167密码子与κ-CnA对应。牦牛中κ-CnG的终止密码子与该基因其他等位基因的终止密码子不同:为TGA,而非TAA。κ-CnH外显子4的核苷酸序列与牛κ-CnA有15个核苷酸替换,导致蛋白质序列中有10个氨基酸变化,这与水牛中κ-酪蛋白相应的已知氨基酸序列一致。揭示了所研究物种等位基因频率分布的品种间和种间差异。讨论了与κ-酪蛋白等位基因进化相关的问题。