大熊猫核糖体蛋白S15A亚基基因(rps15A)的克隆及序列分析  

Cloning and sequence analysis of ribosomal protein S15A(rps15A) gene from the giant panda(Ailuropoda melanoleuca)

作　　者：李俊[1] 丁祥[1] 侯怡铃[1] 孙冰[1] 苏秀兰[1] 吴光富[1] 宋嫣[1] 侯万儒[1] 

机构地区：[1]西华师范大学生命科学学院,南充637002

出　　处：《四川大学学报（自然科学版）》2011年第6期1456-1462,共7页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(30470261);四川省应用基础项目(2009JY0061);四川省教育厅青年基金项目(0928088)

摘　　要：根据已报道的部分物种的核糖体蛋白S15A亚基基因(rps15A)的相关信息设计引物,运用RT-PCR和Touchdown-PCR技术,分别克隆了大熊猫核糖体蛋白S15A亚基的cDNA和结构基因序列,并进行测序及分析.结果表明:大熊猫核糖体蛋白S15A亚基基因的表达序列全长为460 bp,开放阅读框(ORF)为393 bp,编码130个氨基酸,结构基因序列全长为6091 bp,含有4个外显子和3个内含子.RPS15A蛋白的相对分子质量为14.84 kD,pI为10.62.拓扑预测显示含有5个类型的功能位点,即:1个依赖cAMP和cGMP的蛋白激酶磷酸化位点,2个蛋白激酶C磷酸化位点,1个乙酰化位点,1个N-糖基化位点及1个核糖体蛋白S8 signature位点.进一步对RPS15A蛋白的结构分析及其与部分脊椎动物和果蝇RPS15A蛋白的同源性分析,发现该基因的表达序列及其编码的氨基酸序列具有很高的相似性,这表明真核生物核糖体蛋白亚基S15A在进化中具有较高的保守性.The RT-PCR and Touchdown-PCR strategy were employed to isolate cDNA and genome of rpsl5A from the Giant Panda, respectively, sequenced and analyzed. The result showed that the cDNA fragment cloned was 460 bp length, containing an open reading frame（ORF）of 393 bp. The protein deduced was composed of 130 amino acids with an estimated molecular weight of 14. 84 kD and pI of 10.62. The length of the genomic sequence is 6091 bp, which was found to possess 4 exons and 3 in- trons. There are 5 different patterns of functional sites, one N-glycosylation site,one cAMP and cGMP dependent protein kinase phosphorylation sites, two protein kinase C phosphorylation sites, one Amidation site,one Ribosomal protein $8 signature in the Giant Panda RPS15A protein. The deduced amino acid sequence of rps15A was analyzed and compared with human, other vertebrate, invertebrate of rps15A. The results showed that rps15A was highly similarity in eight other species： Homo sapiens, Bos Taurus, Pongo abelii, Rattus norvegicus, Mus musculus, Xenopus tropicalis, Danio rerio, Drosophila melanogaster, suggesting RPS15A protein was conserved highly in evolution.

关 键 词：大熊猫 rps15A 克隆 序列分析 

分 类 号：Q751[生物学—分子生物学]