山羊IGFBP-1基因的克隆及其mRNA丰度在多种组织中的发育性变化  

Molecular Cloning and mRNA Developmental Expression of IGFBP-1 Gene in Goat Different Tissues

作　　者：李利[1,2] 李秋[1] 王林杰[1] 张红平[1] 杜立新[2] 

机构地区：[1]四川农业大学动物遗传育种研究所,雅安625014 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《畜牧兽医学报》2011年第7期899-904,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省教育厅重点项目(08ZA077);四川省科技创新产业链示范工程项目(2011NZ0003)

摘　　要：本研究旨在探讨胰岛素样生长因子结合蛋白1(Insulin-like growth factor-binding protein 1,IGFBP-1)基因在山羊出生后不同组织的发育性表达模式。在出生后0、15、30、60、90和120d共屠宰36只南江黄羊羔羊(公、母羔羊各18只),取心脏、肝脏、脾脏、肺脏、背最长肌、半膜肌、臂三头肌和股二头肌样品以抽提RNA,进行克隆及荧光定量PCR分析。山羊IGFBP-1基因的ORF(Open reading frame)长792bp,共编码263个氨基酸,其核苷酸和氨基酸序列相似性在反刍动物(牛、绵羊)之间远大于其它物种。羔羊肝脏IGFBP-1mRNA表达量最高(P<0.01);肺脏、脾脏、心脏表达水平中等;肌肉中最低且在4种肌肉间表达差异不显著(P>0.05)。出生后尤其是60d内羔羊IGFBP-1mRNA丰度变化很大,且明显呈现持续下降(肝脏)、先升后降(心脏)以及下降-上升-下降(脾脏、肺脏和肌肉)3种模式的变化。结果提示,IGFBP-1基因CDS区在物种间保守性较强,肝脏是山羊合成IGFBP-1mRNA的主要部位,IGFBP-1基因在出生后羔羊的早期生长中发挥着重要的作用且其mRNA发育性表达模式具有组织器官特异性。The objective of the present study was to investigate the developmental expression patterns of insulin-like growth factor-binding protein 1（IGFBP-1） gene in different tissues of postnatal goat.The samples of heart,liver,spleen,lung,longissimus dorsi,semimembranosus,m.triceps brachii and biceps muscle of thigh were collected from a total of 36（18 males and 18 females,respectively） Nanjiang Mongolian Grazelle at the ages of 0,15,30,60,90 and 120 days after birth.The CDS and ontogeny mRNA levels of IGFBP-1 were obtained and analyzed by real-time fluorescence quantitative RT-PCR.The size of IGFBP-1 ORF was 792 bp encoding 263 amino acid residues,and displayed higher nucleotide/amino acid sequence identities in ruminants compared to others.The IGFBP-1 mRNA levels in liver were most abundant（P〈0.01）,medium in lung,spleen and heart,and the lowest in the muscles and had no significant difference among themselves（P〉0.05）.In addition,three expression patterns of IGFBP-1 mRNA during postnatal growth from birth to day 60 were found： consistent down-expression（liver）,fluctuating expression as up-down（heart） or as down-up-down（spleen,lung and muscles）.These results indicate that IGFBP-1 gene is highly conservative among species,and liver is the main expression site of goat IGFBP-1 mRNA and IGFBP-1 play an important role in early growth of postnatal goat,and it is expressed in a developmental-tissue-dependent manner.

关 键 词：山羊 IGFBP-1基因 分子克隆 实时荧光定量RT-PCR 表达 

分 类 号：S827.2[农业科学—畜牧学]