大熊猫核糖体蛋白亚基RPS7基因的克隆及序列分析  

cDNA cloning and sequence analysis of ribosomal protein S7 gene from giant panda

作　　者：侯怡铃[1] 伍春莲[1] 侯万儒[1] 郝彦哲[1] 张田[1] 

机构地区：[1]西华师范大学生命科学学院,四川南充637002

出　　处：《第三军医大学学报》2009年第18期1740-1743,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30470261);四川省应用技术项目(2006J13-057);四川省教育厅重点科研项目(07ZA120);四川省重点学科动物学建设资助项目(404001);四川省科技厅应用基础项目(2009JY0061)~~

摘　　要：目的了解大熊猫(Ailuropoda melanoleuca)核糖体蛋白亚基RPS7基因的结构及其与已报道的人和其他哺乳动物核糖体蛋白亚基RPS7基因的同源性。方法运用RT-PCR技术,从大熊猫的肌肉组织总RNA中对核糖体蛋白亚基RPS7基因的表达序列进行克隆、测序;采用Gnescan软件,对所克隆的基因序列进行氨基酸序列推定;采用ORF finder软件进行DNA序列的开放阅读框(ORF)查找;采用DNAMAN Version6,对基因序列和氨基酸序列进行同源性比较;采用Ex2PASy Proteomics Server软件进行蛋白质功能位点和生化特性预测分析。结果大熊猫RPS7基因的表达序列全长为589bp,ORF为585bp,编码194个氨基酸,该蛋白的相对分子质量为22.12685×103,等电点为10.09,含有1个N-糖基化位点,2个依赖于cAMP和cGMP的蛋白激酶磷酸化位点,4个蛋白激酶C磷酸化位点,1个酪蛋白激酶Ⅱ磷酸化位点,2个十四(烷)酰化位点,2个酰胺化位点及1个RPS7蛋白signature位点。进一步分析结果显示,大熊猫RPS7基因的表达序列及其编码的氨基酸序列与已报道的部分哺乳动物具有很高的相似性。结论运用分子生物学原理与相应的技术手段,成功地扩增出大熊猫RPS7基因的表达序列,并对其编码的蛋白进行了初步分析,丰富和完善了哺乳动物RPS7基因资料库。Objective To explore the structure of ribosomal protein S7 （RPS7） gene of giant panda （Ailuropoda melanoleuca） and investigate its homologies with other already reported sequences, including Homo sapiens, Mus musculus, Rattus norvegicus and Bos taurus. Methods The cDNA of RPS7 was cloned from the giant panda by RT-PCR. The sequence data were analyzed by GenScan software. Blast 2.1 was used to study the homology of the obtained RPS7 sequence with the gene sequences of other species; Open reading frame （ORF） of the DNA sequence was searched using ORF finder software; Protein structure of the RPS7 sequence cloned was deduced using Predict Protein software. Results The full length of the sequence fragment was 589 bp containing an ORF of 585 bp. The deduced protein sequence showed that the protein was composed of 194 amino acids and its estimated molecular weight was 22. 126 85 × 10^3 with a pI of 10.09. There were 7 different patterns of functional sites： one N-glycosylation site; two cAMP and cGMP-dependent kinase phosphorylation sites; four casein kinase C phosphorylation sites; one casein kinase 11 phosphorylation site; two N-myristoylation sites; two amidation sites and one ribosomal protein STe signature site in the RPS7 protein. Further analysis indicated that the sequence of RPS7 and the protein encoded were highly homologous to some mammals reported.Conclusion The complete coding sequence of RPS7 gene has been cloned through RT-PCR successfully, which is the first report on the RPS7 gene from the giant panda.

关 键 词：大熊猫 RT—PCR RPS7 克隆 序列分析 

分 类 号：Q513.1[生物学—生物化学] Q754