野猪骨桥蛋白基因表达及与家猪的对比分析  

Expression of Osteopontin Gene in Sus scrofa and its Comparison with Swine

作　　者：张冬杰[1] 刘娣 杨国伟[1] 汪晓鸿[1] 孙丽华[2] 

机构地区：[1]黑龙江省农业科学院,哈尔滨150086 [2]华南理工大学生物科学与工程学院,广州510640

出　　处：《动物学杂志》2009年第2期41-47,共7页Chinese Journal of Zoology

基　　金：黑龙江省博士后基金项目(No.LRB07-392)

摘　　要：采用RT-PCR法克隆野猪(Sus scrofa)OPN基因,构建其进化树,半定量RT-PCR法结合Western-blot法对其表达进行研究。结果表明,野猪的OPN基因CDS全长909 bp,编码303个氨基酸。特征基序包括(1)第86个氨基酸处有8个连续的天冬氨酸;(2)第153～157氨基酸处有一个与细胞黏附有关的Gly-Arg-Gly-Asp-Ser(GRGDS)序列;(3)第163～164氨基酸处有一个凝血酶酶切位点RS。分子进化树表明,野猪与家猪的亲缘关系最近,与石斑鱼(Danio rerio)的亲缘关系最远,OPN基因在进化过程中发生过一次大的变异。OPN mRNA在心、胃、肾、卵巢的表达水平较高,在肝、脾、肺、小肠、肌肉、子宫内的表达水平较低。不同组织表达的OPN蛋白大小不同,存在70 ku、70 ku+45 ku和70 ku+45 ku+24 ku这三种模式。Sus scrofa OPN gene was cloned by RT-PCR, its phylogenetic tree was constructed, and its expression was studied by semi-quantity RT-PCR combined Western-blot. The result showed that complete CDS of OPN was 909 bp, coded 303 amino acids, and its characteristic motifs included： （1） a potential calcium phosphate apatite binding region of eight consecutive Asp residues beginning at amino acid 86; （2） a cell-attachment Gly-Arg-Gly-Asp-Ser （GRGDS） sequence （residues 153- 157）; and （3） a thrombin RS cleavage site （residues 163 and 164）. The phylogenetic tree showed that the relationship between S. scrofa and swine was the most close, but farthest between S. scrofa and Danio rerio. OPN gene experienced a wide variation during evolution. S. scrofa OPN mRNA was extensively expressed in different tissues, higher in heart, stomach, kidney, ovary, and lower in liver, spleen, lung, small intestine, muscle, and uterus. Different tissues expressed proteins with different weights and there were three patterns of expression, i.e. , 70 ku, 70 ku and 45 ku, as well as 70 ku, 45 ku and 24 ku.

关 键 词：野猪 OPN RT—PCR 分子进化树 Western—blot 

分 类 号：Q953[生物学—动物学]