内蒙古白绒山羊翻译控制肿瘤蛋白(TCTP)基因克隆及组织表达特性分析  

Cloning and Tissue-specific Expression of TCTP Gene in Inner Mongolia Cashmere Goat(Capra hircus)

作　　者：杨立敏[1] 姚睿原 郭志新[1] 朝格图[1] 其布日[1] 郑旭[1] 鲍文蕾[1] 王志钢[1] 

机构地区：[1]内蒙古大学生命科学学院,呼和浩特010021

出　　处：《生物技术通报》2014年第7期125-130,共6页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31160469)

摘　　要：旨在克隆内蒙古白绒山羊翻译控制肿瘤蛋白(Translationally controlled tumor protein,TCTP)基因并分析其表达模式。采用RT-PCR技术扩增TCTP基因编码区cDNA序列,将得到的基因cDNA序列及其编码的氨基酸序列进行生物信息学分析,利用定量RT-PCR方法检测TCTP基因在绒山羊不同组织中的表达特异性。获得的内蒙古白绒山羊TCTP基因编码区cDNA序列全长519 bp,包含了完整的ORF,编码172个氨基酸残基组成的蛋白质。核苷酸序列与绵羊、牛、猪、人、猴及大鼠的同源性在99%-95%之间。生物信息学分析表明,编码的蛋白质理论分子质量19.6 kD,等电点(pI)4.673,含有一个N端糖基化位点,一个蛋白激酶C磷酸化位点,3个酪蛋白激酶Ⅱ磷酸化位点,定位于细胞质中。定量RT-PCR方法检测表明,TCTP基因在绒山羊肾脏、肌肉、胰腺、肝脏、睾丸和脑组织中均有表达,其中在肝脏中的表达量较高,在脑中表达量较低。The present study aims at cloning the CDS fragment of Translationally controlled tumor protein（TCTP）gene cDNA in Inner Mongolia Cashmere Goat and analyzing its expression pattern. TCTP gene cDNA was cloned by RT-PCR. The nucleotide sequence was analyzed by BLAST, while amino acid sequence was analyzed by online softwares. The tissue-specific expression pattern of TCTP was analyzed by quantitative RT-PCR. The cloned TCTP gene cDNA was 519 bp in length, including a complete ORF encoding 172 amino acids. The full cDNA nucleotide sequence has an identity from 99%to 95%with Ovis aries, Bos taurus, Sus scrofa, Homo sapiens, Macaca mulatta and Rattus norvegicus. Bioinformatics analysis showed that theoretical molecular mass of encoded protein was 19.6 kD, isoelectric point（pI）was 4.673. It contained a N-glycosylation sites, a protein kinase C phosphorylation sites and three casein kinaseⅡphosphorylation sites and most probably is localized in cytoplasm. The results of quantitative RT-PCR showed that TCTP gene is expressed in kidney, muscle, pancreas, liver, testis and brain. The expression was higher in liver, whereas lower in brain.

关 键 词：内蒙古白绒山羊 翻译控制肿瘤蛋白 基因克隆 表达模式 

分 类 号：S827[农业科学—畜牧学]