野猪骨桥蛋白基因表达及与家猪的对比分析

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.

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

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...