苏钟猪FTO基因的mRNA表达、克隆测序及其生物信息学分析  

The mRNA Expression,Cloning and Sequencing of FTO gene and Bioinformatics Analysis in Suzhong Pig

作　　者：付言峰[1] 李兰[2,3] 王学敏[1] 李碧侠[1] 方晓敏[1] 赵芳[1] 任守文[1] 

机构地区：[1]江苏省农业科学院畜牧研究所,江苏南京210014 [2]中国农业大学动物医学院,北京100193 [3]国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《华北农学报》2013年第1期128-134,共7页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金(31201767);江苏省博士后科研资助计划经费;江苏省农业科学院博士后基金(No.026056511104);国家生猪现代产业技术体系南京综合试验站(nycytx-009)

摘　　要：脂肪肥胖相关基因(FTO)在控制食欲和能量消耗方面具有重要作用。为进一步了解猪FTO基因的结构和功能,以苏钟猪为试验对象,利用RT-PCR方法,分析该基因在苏钟猪5种不同组织(肺脏、肝脏、心脏、背膘和背最长肌)RNA水平的表达谱信息;对苏钟猪FTO基因的编码序列(CDS)进行了克隆测序,并用生物信息学方法分析了苏钟猪FTO蛋白的结构和不同物种间的同源性。结果表明,FTO基因在苏钟猪各组织中的mRNA表达丰度表现为:背膘>心脏>肺脏>肝脏>背最长肌,其中背膘中的表达量显著高于其他组织(P<0.05)。苏钟猪FTO的CDS区发现了8个SNPs,分别为G52A、C523T、G628A、A655G、A810G、A886G、G1002A和A1344G。苏钟猪FTO基因编码一个含505个氨基酸的蛋白,理论等电点为5.23,具有亲水性,不存在信号肽,是非分泌型蛋白,与牛、绵羊、狗等物种的亲缘关系最近。FTO的mRNA表达对苏钟猪的皮下脂肪、心脏、肌肉等组织的脂肪沉积有一定影响,其编码序列中丰富的SNPs可成为苏钟猪肉质改良育种中的重要分子标记。Fat mass and obesity associated gene（FTO） plays an important role in the control of appetite and energy expenditure.In order to understand the structure and function of porcine FTO gene,the mRNA expression profile of porcine FTO in five tissues（lung,liver,heart,backfat and longissimus dorsi muscle） were examined in Suzhong pigs by RT-PCR.Besides,the coding sequences（CDS） of FTO were cloned and sequenced in Suzhong pigs,molecular structure and homology among different species of FTO were analyzed by bioinformatics methods.The results showed that mRNA expression intensity of FTO in different tissues tended to be backfatheartlungliverlongissimus dorsi muscle.Among these tissues,FTO expression in backfat was significantly higher than that in any other tissue（P0.05）.There were eight SNPs detected in coding sequences（CDS） of FTO in Suzhong pigs,including G52A,C523T,G628A,A655G,A810G,A886G,G1002A and A1344G.Bioinformatic analysis revealed that porcine FTO gene encode a protein of 505 amino acid,and its theoretical isoelectric point was 5.23.FTO protein was a hydrophilic and non-secreted protein without a signal peptide.In addition,Suzhong pig was closed to cattle,sheep and dog in phylogenetic tree.These findings suggest that the mRNA expression of FTO may have certain influence in regulating fat deposition in some tissues of Suzhong pigs,such as subcutaneous fat,heart and muscle,etc,and these abundant SNPs in coding sequence of FTO could become important molecular markers for improving meat quality in Suzhong pigs.

关 键 词：苏钟猪 脂肪肥胖相关基因 克隆测序 遗传变异 生物信息学 

分 类 号：Q78[生物学—分子生物学]