利用PCR-RFLP和PCR-SSCP技术检测泽羊肌肉生长抑制素基因第1和2内含子及第3外显子的多态性及其与周岁体重的关联
Polymorphism of Myostatin Gene in Intron 1 and 2 and Exon 3, and Their Associations with Yearling Weight, Using PCR-RFLP and PCR-SSCP Techniques in Zel Sheep.
作者信息
Dehnavi Elena, Ahani Azari Mojtaba, Hasani Saeed, Nassiry Mohammad Reza, Mohajer Mokhtar, Khan Ahmadi Alireza, Shahmohamadi Leila, Yousefi Soheil
机构信息
Department of Animal Science, Gorgan University of Agricultural Sciences and Natural Resources, P.O. Box 4913815739, Gorgan, Iran.
出版信息
Biotechnol Res Int. 2012;2012:472307. doi: 10.1155/2012/472307. Epub 2012 Jun 20.
The aim of present study was to investigate myostatin gene polymorphism and its association with yearling weight records in Zel sheep using PCR-RFLP and PCR-SSCP methods. Blood samples were collected from 200 Zel sheep, randomly, and DNA was extracted using modified salting out method. Polymerase chain reaction was carried out to amplify 337, 222, and 311 bp fragments, respectively, comprising a part of exon 3, intron 1, and intron 2 of myostatin gene. In addition, exon 3 was digested by HaeIII enzyme under RFLP method, and introns 1 and 2 were studied using SSCP. Under RFLP method, all samples showed mm genotype. Under SSCP method, intron 1 was also monomorph but intron 2 was polymorph (AA, AB, and BB). The allelic frequencies for A and B were 75.5 and 24.5%, respectively. This locus was not in Hardy-Weinberg equilibrium (P < 0.05), and there was no significant effect of myostatin gene on yearling weights.
本研究旨在利用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）和聚合酶链式反应-单链构象多态性（PCR-SSCP）方法，研究泽羊肌生成抑制素基因多态性及其与周岁体重记录的关联。随机采集200只泽羊的血样，采用改良盐析法提取DNA。进行聚合酶链式反应，分别扩增肌生成抑制素基因外显子3、内含子1和内含子2的一部分，片段长度分别为337、222和311 bp。此外，在RFLP方法下，用HaeIII酶消化外显子3，利用SSCP研究内含子1和内含子2。在RFLP方法下，所有样本均显示为mm基因型。在SSCP方法下，内含子1也是单态的，但内含子2是多态的（AA、AB和BB）。A和B的等位基因频率分别为75.5%和24.5%。该位点不符合哈迪-温伯格平衡（P < 0.05），肌生成抑制素基因对周岁体重没有显著影响。
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