黑尾足鼠核糖体蛋白S18基因(RPS18)的克隆及序列分析

Xiao Cui, Zheng-song Peng, Jun Yang, Yi-ling Hou, Xiang Ding, Wan-Ruhou
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摘要

核糖体蛋白S18是真核细胞中由RPS18基因编码的40S亚基的一个组成部分,属于核糖体蛋白S13P家族。为探究黑尾足鼠RPS18基因的结构特征,我们根据已知的RPS18基因核苷酸序列重新设计引物,分别采用RT-PCR和PCR策略克隆黑尾足鼠RPS18基因cDNA和基因组序列,并对克隆的cDNA和基因组序列进行初步测序和分析。结果表明,大熊猫RPS18基因cDNA片段全长503 bp,包含459 bp的开放阅读框(ORF),编码152个氨基酸。基因组序列长度为5518 bp,包含6个外显子和5个内含子。编码序列与智人、金牛、小家鼠、scrofa、狼犬和褐家鼠具有高度的同源性;同源度分别为92.16%、92.59%、86.64%、91.50%、94.77%和89.54%。而所有氨基酸序列的同源性都很高,高达98.68%。初步结构分析表明,推测的RPS18蛋白分子量为17.65 KD,理论pI为11.39。基于拓扑预测,大熊猫RPS18蛋白存在1个酪氨酸激酶磷酸化位点、3个蛋白激酶C磷酸化位点、2个酪蛋白激酶II磷酸化位点、3个酰胺化位点和1个核糖体蛋白S13特征。
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Cloning and sequence analysis of ribosomal protein S18 gene (RPS18) from Ailuropoda melanoleuca
Ribosomal protein S18 is a component of the 40S subunit, encoded by the RPS18 gene in eucaryotic cell, belonging to the S13P family of ribosomal proteins. To explore the structure characteristic of RPS18 gene of the Ailuropoda melanoleuca, primers we re designed based on the known nucleotide sequence of RPS18 gene to clone the cDNA and genomic sequences of RPS18 gene of A. melanoleuca by RT-PCR and PCR strategy respectively, and then the cloned cDNA and genomic sequences were sequenced and analyzed preliminarily. The results indicated that the length of cDNA fragment of the RPS18 from the giant panda is 503 bp in size, containing an open reading frame (ORF) of 459 bp, encoding 152 amino acids. The length of the genomic sequence is 5,518 bp, with six exons and five introns. The coding sequence shows a high degree of homology to those of Homo sapiens, Bos taurus, Mus musculus, Sus scrofa, Canis lupus and Rattus norvegicus; the degree of homology was 92.16 %,92.59%, 86.64%, 91.50%, 94.77% and 89.54%, respectively. While all the homologies for amino acid sequences are high up to 98.68%. Primary structure analysis revealed that the molecular weight of the putative RPS18 protein is 17.65 KD, with a theoretical pI of 11.39. Based on topology prediction, there is one Tyrosine kinase phosphorylation site, three protein kinase C phosphorylation site, two casein kinase II phosphorylation site, three amidation site and one ribosomal protein S13 signature in the RPS18 protein of the giant panda.
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