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首页> 外文期刊>Genetics and Molecular Research >Giant panda ribosomal protein S14: cDNA, genomic sequence cloning, sequence analysis, and overexpression
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Giant panda ribosomal protein S14: cDNA, genomic sequence cloning, sequence analysis, and overexpression

机译：大熊猫核糖体蛋白S14：cDNA，基因组序列克隆，序列分析和过表达

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RPS14 is a component of the 40S ribosomal subunit encoded by the RPS14 gene and is required for its maturation. The cDNA and the genomic sequence of RPS14 were cloned successfully from the giant panda (Ailuropoda melanoleuca) using RT-PCR technology and touchdown-PCR, respectively; they were both sequenced and analyzed. The length of the cloned cDNA fragment was 492 bp; it contained an open-reading frame of 456 bp, encoding 151 amino acids. The length of the genomic sequence is 3421 bp; it contains four exons and three introns. Alignment analysis indicates that the nucleotide sequence shares a high degree of homology with those of Homo sapiens, Bos taurus, Mus musculus, Rattus norvegicus, Gallus gallus, Xenopus laevis, and Danio rerio (93.64, 83.37, 92.54, 91.89, 87.28, 84.21, and 84.87%, respectively). Comparison of the deduced amino acid sequences of the giant panda with those of these other species revealed that the RPS14 of giant panda is highly homologous with those of B. taurus, R. norvegicus and D. rerio (85.99, 99.34 and 99.34%, respectively), and is 100% identical with the others. This degree of conservation of RPS14 suggests evolutionary selection. Topology prediction shows that there are two N-glycosylation sites, three protein kinase C phosphorylation sites, two casein kinase II phosphorylation sites, four N-myristoylation sites, two amidation sites, and one ribosomal protein S11 signature in the RPS14 protein of the giant panda. The RPS14 gene can be readily expressed in Escherichia coli. When it was fused with the N-terminally His-tagged protein, it gave rise to accumulation of an expected 22-kDa polypeptide, in good agreement with the predicted molecular weight. The expression product obtained can be purified for studies of its function.
机译：RPS14是RPS14基因编码的40S核糖体亚基的组成部分，是其成熟所必需的。分别使用RT-PCR技术和触地PCR技术成功地从大熊猫中克隆了RPS14的cDNA和基因组序列。他们都进行了测序和分析。克隆的cDNA片段的长度为492bp。它包含一个456 bp的开放阅读框，编码151个氨基酸。基因组序列的长度为3421 bp;它包含四个外显子和三个内含子。比对分析表明该核苷酸序列与智人，金牛座，小家鼠，褐家鼠，盖氏鸡，非洲爪蟾和雷氏达尼奥（93.64、83.37、92.54、91.89、87.28、84.21，和84.87％）。比较大熊猫与其他物种的推导氨基酸序列后发现，大熊猫的RPS14与金牛座牛，诺维发牛和雷氏暴走牛的RPS14高度同源（分别为85.99％，99.34％和99.34％） ），并且与其他100％相同。 RPS14的这种保守程度表明进化选择。拓扑预测表明，大熊猫的RPS14蛋白中有两个N-糖基化位点，三个蛋白激酶C磷酸化位点，两个酪蛋白激酶II磷酸化位点，四个N-肉豆蔻酰基化位点，两个酰胺化位点和一个核糖体蛋白S11签名。 。 RPS14基因很容易在大肠杆菌中表达。当其与N末端His标签的蛋白融合时，它引起了预期的22kDa多肽的积累，与预测的分子量非常一致。可以纯化获得的表达产物以研究其功能。

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