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Cloning and comparative analysis of gene structure in promoter site of alpha-s1 casein gene in Naeinian goat and sheep

机译:奈爱尼亚山羊和绵羊α-s1酪蛋白基因启动子位点基因结构的克隆和比较分析

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摘要

The 5′ end or alpha-S1 casein promoter has a significant role in milk protein gene expression. The understanding of the translation process of alpha-S1 casein mutants will provide us an opportunity to make the best selection in livestock providing more proteins in milk. Blood samples were taken from three hundred of Naeinian goats and sheep, and DNA extraction was done using modified salting out method. Polymerase chain reactions (PCR) were carried out using a specific primer pairs for amplification a fragment of 1133 bp from part of 5′-UTR and exon 1 of alpha s1 casein gene. The AluI and HinfI restriction enzyme treatment of all samples provided the same homozygous AA genotype in both species. Subsequently, one sample of each species was selected and cloned, and the final sequences were analyzed by BioEdit, CLC genomic, Mega4 and DNASIS MAX software. Several polymorphisms are recognized between Naeinian goat and sheep that are presented on motif sites. In this research, the interested location, including exon I and a part of 5′, was analyzed, and genetic element comparisons were done between Naeinian goat and sheep. The number and location of probable binding sites can have a crucial role as a result of antagonistic and synergistic effects on gene regulation activities.
机译:5'端或α-S1酪蛋白启动子在乳蛋白基因表达中具有重要作用。对α-S1酪蛋白突变体翻译过程的理解将为我们提供机会,使家畜中的最佳选择提供更多的牛奶中的蛋白质。从三百只奈伊尼亚山羊和绵羊中采集血样,并使用改良的盐析法提取DNA。使用特定引物对进行聚合酶链反应(PCR),以扩增5'-UTR的一部分和alpha s1酪蛋白基因外显子1的1133bp片段。所有样品的AluI和HinfI限制酶处理在两个物种中提供了相同的纯合AA基因型。随后,选择并克隆每个物种的一个样本,并通过BioEdit,CLC基因组,Mega4和DNASIS MAX软件分析最终序列。奈伊尼亚山羊和绵羊之间在母题位点上发现了几种多态性。在这项研究中,分析了感兴趣的位置,包括外显子I和5'的一部分,并对Naeinian山羊和绵羊进行了遗传元素比较。可能的结合位点的数量和位置可能起关键作用,这是对基因调节活性产生拮抗和协同作用的结果。

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