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纳尼山羊和绵羊α-s1酪蛋白基因启动子区基因结构的克隆与比较分析

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

作者信息

Najafi Mojtaba, Rahimi Mianji Ghodrat, Ansari Pirsaraie Zarbakht

机构信息

Department of Animal Science, Sari Agriculture sciences and Natural Resources University, Iran.

出版信息

Meta Gene. 2014 Nov 16;2:854-61. doi: 10.1016/j.mgene.2014.11.001. eCollection 2014 Dec.

Abstract

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.

摘要

5'端或α-S1酪蛋白启动子在乳蛋白基因表达中起着重要作用。对α-S1酪蛋白突变体翻译过程的了解将为我们提供一个机会,以便在产奶中能提供更多蛋白质的家畜中做出最佳选择。从三百只纳尼羊和山羊采集血样,采用改良盐析法进行DNA提取。使用特异性引物对进行聚合酶链反应(PCR),以扩增α-s1酪蛋白基因5'-UTR部分和外显子1的1133 bp片段。对所有样本进行AluI和HinfI限制性酶切处理,结果表明这两个物种均为相同的纯合AA基因型。随后,从每个物种中选取一个样本进行克隆,并使用BioEdit、CLC genomic、Mega4和DNASIS MAX软件对最终序列进行分析。在纳尼羊和山羊之间识别出了几个存在于基序位点的多态性。在本研究中,对包括外显子I和5'端一部分在内的感兴趣区域进行了分析,并对纳尼羊和山羊之间的遗传元件进行了比较。由于对基因调控活动的拮抗和协同作用,可能的结合位点的数量和位置可能起着关键作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b03f/4287881/98b1465e0f69/gr1.jpg

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