绵羊抵抗素基因(RETN)克隆、序列分析及其在饥饿模型中的表达变化规律  

Cloning and Sequence Analysis of Sheep(Ovis aires) Resistin Gene(RETN) and Its Expression Pattern in Starving Model

作　　者：赵伟利[1,2] 薛昌斌[3] 高磊[2] 许瑞霞[1,2] 宋广超[1,2] 张伟[1,2] 刘守仁[2] 王新华[2] 杨井泉[2] 尹君亮[2] 甘尚权[2] 

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]新疆农垦科学院畜牧兽医研究所/省部共建绵羊遗传改良与健康重点实验室,石河子832000 [3]新疆生产建设兵团第十二师三坪农场,乌鲁木齐830000

出　　处：《农业生物技术学报》2015年第8期981-990,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金新疆联合基金重点项目(No.U1130302);国家基础研究发展计划(973)前期专项项目(No.2011CB111501);新疆农垦科学院引导计划(No.YYD2010-9);兵团国际科技合作计划(No.2012BC003)

摘　　要：抵抗素(resistin,RETN)是脂肪细胞特异性分泌的激素,其在葡萄糖与脂类代谢中发挥重要作用,为研究RETN基因在绵羊(Ovis aires)尾脂沉积与代谢中的表达变化规律,本研究采用长片段PCR技术首次克隆获得绵羊RETN基因(Gen Bank登录号:KJ704841)全长序列。生物信息学分析表明,RETN基因编码有别于已公布的绵羊RETN序列,编码109个氨基酸;利用半定量RT-PCR技术检测RETN基因组织表达谱,结果表明,RETN基因在绵羊肝、脾、肺及臀脂等组织中均有不同程度的表达,肝脏中的表达丰度最高,极显著高于其他组织(P<0.01),暗示RETN基因在脂肪合成与代谢重要器官肝脏中可能发挥着重要生理生化功能;采用构建的阿勒泰羊持续饥饿模型实时荧光定量PCR(fluorescence quantitative Realtime PCR,q RT-PCR)分析阿勒泰羊在饥饿与否状态下臀脂的表达差异,结果显示,RETN m RNA表达在两种极端条件下臀脂中差异极显著(P<0.01),持续饥饿状态下RETN基因表达量是正常组的近5.2倍,提示RETN基因可能在脂肪组织的动员过程中发挥重要作用。以上研究结果为进一步揭示RETN基因在绵羊臀脂中的生物功能提供基础数据。Resistin （RETN） is a specifically produced hormone by fat cells, which plays an important role in glucose and lipid metabolism and also is one of popular candidate genes in animal fat deposition and metabolism. Aiming at study the expression pattern of RETN gene in rump-fat deposition and metabolism of sheep （Ovis aires）, long fragment PCR, bioinformatics method and semi-quantitative RT-PCR were used toamplify the full-length sequences, analyze the sequences and detect gene expression profiling of sheep RETN gene, respectively. Fluorescence quantitative Real-time PCR （qRT-PCR） method was used to analyze the expression change of RETN gene between starving and its control groups based on starving model in Altay sheep. The results of cloning and sequence analysis showed that RETN gene encoded 109 amino acids （GenBank No. KJ704841）, which was different from the published RETN sequence in GenBank database. RETN mRNA expression profiling showed that RETN expressed in liver, spleen, lung and rump fat differently, especially highly in the liver, significantly higher than that of other groups （P〈0.01）, which indicated that RETN gene might play important physiological and biochemical functions in fat synthesis and metabolism in the liver. The result of qRT-PCR showed that RETN mRNA significantly differentially expressed in between starving group and its control groups, with a expression level in persistent starvation group of 5.2 times than that of control group, suggesting that RETN gene might play an important role in fat mobilization. The above findings provide fundamental data for further study on RETN biological function in fat-rump sheep.

关 键 词：抵抗素(RETN) 饥饿模型 臀脂 序列分析 表达谱研究 

分 类 号：S852.659.6[农业科学—基础兽医学]