利用PCR-RFLP和PCR-SSCP技术分析泽尔羊1、2内含子和3外显子肌生长抑制素基因多态性及其与初生体重的关系

Biotechnology Research International Pub Date : 2012-01-01 Epub Date: 2012-06-20 DOI:10.1155/2012/472307
Elena Dehnavi, Mojtaba Ahani Azari, Saeed Hasani, Mohammad Reza Nassiry, Mokhtar Mohajer, Alireza Khan Ahmadi, Leila Shahmohamadi, Soheil Yousefi
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引用次数: 26

摘要

采用PCR-RFLP和PCR-SSCP方法研究泽尔羊肌肉生长抑制素基因多态性及其与初生体重的关系。随机抽取200只Zel羊的血液样本,采用改良盐析法提取DNA。聚合酶链反应分别扩增了337、222和311 bp的片段,这些片段包含肌生长抑制素基因外显子3、内含子1和内含子2的一部分。此外,采用RFLP法对HaeIII酶切除外显子3,采用SSCP法对内含子1和2进行了研究。在RFLP法下,所有样品均为mm基因型。SSCP法下,内含子1为单态,内含子2为多态(AA、AB、BB)。A和B的等位基因频率分别为75.5%和24.5%。该基因位点不处于Hardy-Weinberg平衡状态(P < 0.05),肌生长抑制素基因对仔猪出生体重无显著影响。
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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.

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.

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