大熊猫核糖体蛋白S11亚基基因(RPS11)cDNA的克隆及序列分析

cDNA Cloning and Sequence Analysis of the Ribosomal Protein S11 Gene(RPS11)from the Giant Panda

RPS11是核糖体小亚基40S的组成部分,由RPS11基因所编码,属于核糖体蛋白S17p家族,主要存在于真核生物中。为了解大熊猫核糖体蛋白亚基RPS11基因的结构特点及其与已报道的人和其他哺乳动物核糖体蛋白亚基RPS11基因的异同,本研究根据已报道的部分哺乳动物核糖体蛋白S11亚基基因(RPS11)的相关信息设计引物,运用RT-PCR技术从大熊猫的肌肉组织总RNA中成功克隆了核糖体蛋白亚基RPS11基因,并进行了测序和序列分析。结果表明:大熊猫RPS11亚基基因的开放阅读框(ORF)长为477bp,编码158个氨基酸的蛋白质,该蛋白的相对分子量为18.4275kDa,pI为10.96。拓扑预测显示该蛋白含有14个功能位点:即2个N-糖基化位点,6个蛋白激酶C磷酸化位点,4个酪蛋白激酶Ⅱ磷酸化位点,1个酪氨酸激酶磷酸化位点和1个核糖体蛋白S17 signature位点。进一步分析发现,大熊猫RPS11基因与已报道的部分哺乳动物的表达序列及其编码的氨基酸序列都具有很高的相似性。本研究结果为丰富和完善哺乳动物RPS11基因资源库提供了基础资料。

RPS11 is a component of the 40S small ribosomal subunit encoded by the RPS11 gene and belongs to the S17p family of ribosomal proteins,which is specific to eukaryotes.The aim of this study is to explore the structural characteristics of the ribosomal protein S11（RPS11）gene from the giant panda（Ailuropoda melanoleuca）and to investigate its similarities and differences with other reported sequences,including Homo sapiens,Mus musculus,Rattus norvegicus,Bos taurus,Danio rerio and Drosophila melanogaster.Primers used in this study were designed based on information from the ribosomal protein s11 gene（RPS11）from other mammals.The RPS11 was cloned successfully for the first time from the giant panda using RT-PCR technology,its sequence was also sequenced,and preliminary analyses conducted.The open-reading frame（ORF）of 477 bp encoded 158 amino acids.Primary structural analysis revealed that the molecular weight of the putative RPS11 protein was 18.4275 kDa with a theoretical pI of 10.96.Topology prediction showed that there were two N-glycosylation sites,six protein kinase C phosphorylation sites,four casein kinase Ⅱ phosphorylation sites,one tyrosine kinase phosphorylation site,and one ribosomal protein S17 signature in the RPS11 protein of the giant panda.Alignment analysis indicated that the nucleotide sequence and the deduced amino acid sequence shared a high homology with those of mammalians.The cDNA of RPS11 was cloned successfully for the first time from the giant panda in this study.The results of this study provided scientific material for enriching and improving the mammals RPS11 gene database.