梅花鹿茸角组织BSPⅡ基因全长cDNA的克隆及序列特征分析  

Full length cDNA cloning and sequence characteristics analysis of BSP Ⅱ gene from Sika deer antler tissue

作　　者：郝丽[1] 

机构地区：[1]东北林业大学,黑龙江哈尔滨150040

出　　处：《中国兽医学报》2011年第7期1013-1017,1023,共6页Chinese Journal of Veterinary Science

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项目(DL10BA08)

摘　　要：从梅花鹿鹿茸尖端组织全长cDNA文库中克隆了与骨形成和骨改建有关的一种新的骨生长因子BSPⅡ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其在鹿茸尖端不同组织层的表达情况进行了分析。结果表明,BSPⅡ基因cDNA全长为1 576bp,编码311个氨基酸。经生物信息学分析表明,该基因编码的蛋白具有N端信号肽及跨膜区,相对分子质量为34 100,理论等电点为4.05,其一级结构中谷氨酸所占比例最高;二级结构元件主要以α-螺旋和无规则卷曲为主;同源序列分析表明,梅花鹿与欧洲牛BSPⅡ氨基酸的相似性最高(93%);多重序列比对显示,此蛋白的N-端和68～215、265～308位谷氨酸富集区高度保守;系统进化树显示,在该基因座位上梅花鹿与马的亲缘关系较近,来源于同一个分化支。实时荧光定量RT-PCR分析表明,该基因的表达与鹿茸组织的矿化进程呈显著正相关,推测该基因在鹿茸组织矿化过程中起到了重要的调节作用。The full length cDNA of BSP Ⅱ gene from velvet tip tissue full-length cDNA library of Sika deer was cloned,bioinformatics method and Real-time RT-PCR technique were used to analyze the amino acid sequence and expression. The results showed the full-length cDNA of the BSP Ⅱ gene was 1 576 bp,encoded a peptide of 311 amino acid,its relative molecular weight was 34 100, isoelectric point was 4.05, contained a N-terminal signal peptid and transmembrane domain; Glu occupied the highest proportion and secondary structure with a-helix and random coilbased. The results obtained through homologous sequence analysis indicated that BSP Ⅱ of Sika deer was highly sim- ilarity to Bos Taurus（93 %） ;with multiple sequences comparison,the N-end and 68-215,265-308 Glu rich area were highly conservative; the molecular evolution trees displayed that BSP Ⅱ of Sika deer had high genetic relationship with Equus caballus. Real-time PCR results showed that there was a significant positive correlation between its expression and velvet mineralization process showed ,it is presumed that the BSP Ⅱ genes in the process of velvet tissue mineralization plays an important role in regulating.

关 键 词：鹿茸 骨唾液蛋白Ⅱ CDNA文库 基因克隆 

分 类 号：S825[农业科学—畜牧学]