摘要
骨桥蛋白(Osteopontin,OPN)是在骨基质的矿化和吸收过程中起重要作用的细胞因子,研究首次从梅花鹿鹿茸尖端组织cDNA文库中成功克隆了OPN基因的全长cDNA序列,并结合生物信息学方法和实时荧光定量RT-PCR技术对该基因的氨基酸序列及表达特征进行分析。结果表明,OPN的cDNA全长为1 406 bp,编码279个氨基酸。经生物信息学分析,该基因为不稳定蛋白,具有N端信号肽,相对分子质量为30.99 ku,理论等电点为4.40,其一级结构中丝氨酸和谷氨酸残基所占比例最高,含有特异的Arg-Gly-Asp序列(RGD)。同源序列比对及系统进化树分析表明,鹿源OPN氨基酸序列不仅与欧洲牛和山羊相似性最高(分别为86%,85%),且在该基因座位上也与二者在亲缘关系最近。实时荧光定量RT-PCR分析表明,OPN基因在鹿茸尖端不同组织层的表达存在差异,前软骨层和软骨层的表达量普遍高于间充质和皮肤层,推测OPN可能通过介导破骨细胞与矿化组织的黏附,从而参与了鹿茸组织的矿化。
Osteopontin（OPN） is a cell factor which plays an important role in the process of mineralization and absorption of bone matrix.The full length cDNA of OPN from velvet tip tissue full-length cDNA library of Sika Deer was isolated,and bioinformatics method and real-time RT-PCR technique were used to analyze the amino acid sequence and expression.The full-length cDNA of OPN gene was 1 406 bp and encoded a peptide of 279 amino acid residues of which the relative molecular weight was 30.99 ku.By analysis of bioinformatics,the gene had an isoelectric point of 4.40,and contained a N-terminal signal peptid;Ser and Glu occupied the highest proportion and the special sequence Arg-Gly-Asp（RGD） existed in OPN primary structure.The results obtained through homologous sequence analysis indicated that OPN mature protein of Sika Deer was highly similarity with Bos taurus and Capra hircus（86% and 85%）.The molecular evolution trees showed that OPN of Sika Deer had the high genetic relationship with Bos taurus and Capra hircus.Real-time PCR results showed that the gene expression had tissue specificity,and the expression from the skin layer to the precartilage layer was gradually rising while in the cartilage layer expression was descending.It was speculated that the gene was involved in velvet tissue calcification through the osteoclast-mediated adhesion with mineralized tissues.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2010年第12期91-97,共7页
Journal of Northeast Agricultural University
基金
黑龙江省自然科学基金资助项目(C200725)
中央高校基本科研业务费专项资金资助(DL10BA08)
