大白猪骨桥蛋白基因的克隆、表达及功能分析  

Cloning,Expression and Functional Analysis of Osteopontin Gene in Large White Pig

作　　者：张冬杰[1,2] 刘娣 汪晓鸿[2] 杨国伟[2] 

机构地区：[1]东北林业大学博士后流动站,哈尔滨150040 [2]黑龙江省农业科学院畜牧研究所博士后工作分站,哈尔滨150086

出　　处：《畜牧兽医学报》2009年第7期965-970,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：黑龙江省科技攻关项目(GB05B106);黑龙江省博士后基金(LRB07-392)

摘　　要：应用RT-PCR方法克隆了全长909 bp的大白猪Osteopontin(OPN)基因。运用生物信息学工具进行的分析显示:OPN蛋白一级结构中天冬氨酸、谷氨酸、丝氨酸所占比例最高,含有特异的Arg-Gly-Asp序列(RGD),二级结构中α螺旋占很大比例,并且亲水性较强,OPN信号标签位于第一个同源区,共有的保守性序列为[KQ]-x-[TA]-x(2)-[GA]-S-S-E-E-K。基于蛋白序列所构建的2个分子进化树均表明猪与牛的亲缘关系最近,而与鸡最远。在mRNA水平上,OPN基因在猪体各组织中广泛表达,胃、肾、肺、小肠和卵巢相对较高,而心、脾和大肠则相对较低;在蛋白水平上,不同组织内该蛋白的大小不同,出现了70 ku,70和45 ku,70、45和24 ku 3种蛋白。The Osteopontin cDNA in size of 909 bp was cloned by RT-PCR and analyzed by bioinformatics. The results showed that Asp, Glu and Ser occupied the highest proportion and the special sequence Arg-Gly-Asp （RGD） existed in OPN primary structure, a helix occupied the higher proportion in secondary structure and hydrophile was better, OPN signature existed in the first homologous region and the mutual conservative sequence was [KQ]-x-[-TA]-x（2）-[-GA]-S- S-E-E-K in OPN secondary structure. Two phylogentic trees constructed based on OPN protein sequence showed that the relationship between pig and cow was the closest, but distant from chicken. OPN mRNA was expressed in many tissues of pig： higher in stomach, kidney, lung, small intestine and ovary, lower in heart, spleen and large intestine. The protein size was different in different tissues： 70 ku, 70 and 45 ku, 70, 45 and 24 ku.

关 键 词：大白猪 骨桥蛋白基因 生物信息学 反转录PCR 蛋白质斑点杂交 

分 类 号：S828.89[农业科学—畜牧学] Q344.13[农业科学—畜牧兽医]