Genetic Analysis Using Partial Sequencing of Melanocortin 4 Receptor (MC4R) Gene in Bligon Goat

Latifah Latifah, D. A. Priyadi, D. Maharani, K. Kustantinah, T. Hartatik
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引用次数: 7

Abstract

Melanocortin 4 Receptor gene is involved in sympathetic nerve activity, adrenal and thyroid functions, and media for leptin in regulating energy balance and homeostasis. The aim of this research was to perform genetic analysis of MC4R gene sequences from Bligon goats. Fourty blood samples of Bligon does were used for DNA extraction. The primers were designed after alignment of 12 DNA sequences of MC4R gene from goat, sheep, and cattle. The primers were constructed on the Capra hircus MC4R gene sequence from GenBank (accession No. NM_001285591). Two DNA polymorphisms of MC4R were revealed in exon region (g.998 A/G and g.1079 C/T). The SNP g.998 A/G was a non-synonymous polymorphism i.e., changing of amino acid from methionine (Met) to isoleucine (Ile). The SNP g.1079 C/T was a synonymous polymorphism. Restriction enzyme mapping on Bligon goat MC4R gene revealed three restriction enzymes (RsaI (GT’AC), Acc651 (G’GTAC_C), and KpnI (G_GTAC’C), which can recognize the SNP at g.1079 C/T. The restriction enzymes may be used for genotyping of the gene target using PCR-RFLP method in the future research.
黑色素皮质素4受体(MC4R)基因部分测序在Bligon山羊遗传分析中的应用
黑色素皮质素4受体基因参与交感神经活动、肾上腺和甲状腺功能,以及瘦素调节能量平衡和稳态的介质。本研究的目的是对Bligon山羊MC4R基因序列进行遗传分析。使用了四十份Bligon的血液样本进行DNA提取。对山羊、绵羊和牛MC4R基因的12个DNA序列进行比对,设计引物。引物构建在来自GenBank(登录号NM_001285591)的毛山羊MC4R基因序列上。在外显子区发现MC4R的两个DNA多态性(g.998 A/g和g.1079 C/T)。SNP g.998 A/g是一个非同义多态性,即氨基酸从蛋氨酸(Met)变为异亮氨酸(Ile)。SNP g.1079C/T是同义多态性。对Bligon山羊MC4R基因进行限制性内切酶定位,发现三种限制性内切酶RsaI(GT'AC)、Acc651(G'GTAC_C)和KpnI(G_GTAC'C)能识别G.1079C/T的SNP。在未来的研究中,限制性内切酶可用于使用PCR-RFLP方法对基因靶标进行基因分型。
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