仔猪水通道蛋白基因的克隆及断奶应激对其在胃肠道组织mRNA表达的影响  

Cloning and mRNA Expression of Aquaporins Gene in the Gastrointestinal Tract of Weaning Piglets

作　　者：韩立强[1] 台玉磊[1] 郭豫杰[1] 鲁维飞[1] 李宏基[1] 查光明[1] 李奎[2] 杨国宇[1] 

机构地区：[1]河南农业大学农业部动物生化与营养重点开放实验室,郑州450002 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《农业生物技术学报》2012年第8期886-892,共7页Journal of Agricultural Biotechnology

基　　金：国家转基因生物新品种培育重大专项(No.2011ZX08006-003);农业部948重点项目(No.2011-G35)

摘　　要：水通道蛋白(AQP)作为一种膜转运蛋白,对于调控肠道组织的水分子转运具有重要作用。本实验利用同源序列克隆法设计引物,从仔猪(Susscrofa)的消化道组织中克隆AQPs家族基因(AQP2~AQP11)(GenBank登录号分别为EU636238、EU161094、EU165525、EU192130、EU620575、EU024116、EU220426、EU220427、EU582021和EU220425),并对其进行生物信息学分析。采用荧光定量PCR研究断奶应激对于仔猪胃肠道组织中AQP1、AQP3、AQP8和AQP11的mRNA表达影响。生物信息学分析发现,猪AQPs家族基因含有高度保守的天冬酰氨-脯氨酸-丙氨酸(NPA)结构序列,基因表达分析发现,断奶组仔猪的AQP1基因在十二指肠、回肠和结肠组织的mRNA表达量与对照组相比显著上调(P<0.05);AQP3基因在胃和小肠组织的mRNA表达有显著上调(P<0.05);AQP8基因在胃和回肠组织中mRNA表达有显著上调(P<0.01),在其他组织中没有显著差异;AQP11基因在空肠组织的mRNA表达量显著上调了1.7倍(P<0.05)。这些研究结果表明,AQP基因可能在断奶应激引起的仔猪肠道腹泻中发挥重要作用。Aquaporins （AQPs） play importment roles in the water absorbing through gastrointestinal tract as channel proteins. The encoding sequences of aquaporins （A OP2-A OPll） were cloned from the gastrointestinal tract tissue of piglets （Sus scrofa）, and then ran bioinformatics analysis. The GenBank accession numbers were EU636238, EU161094, EU165525, EU192130, EU620575, EU024116, EU220426, EU220427, EU582021 and EU220425, respectively. Eight piglets were divided randomly to control and weaning group. Mucosal tissue was collected from the gastrointestinal tract （stomach, duodenum, jejunum, ileum, colon and rectum） and Real-time PCR was applied to determine the mRNA expression of A QPs genes （A QP1, A QP3, A QP8 and A QPll）. Bioinfbrmatics sot^ware analysis showed that the AQPs had a consevved amino acid sequence of asparagine-proline-alanine （NPA） motif in the protein family. The gene expression results indicated that compared with the control group, mRNA expression ofA QP1 was significantly up-regulated in the duodenum, ileum and colon tissue （17〈0.05）; mRNA expression ofA QP3 was significantly up-regulated in the stomach and small intestine tissue （P〈0.05）; mRNA expression of A QP8 had no significantly difference in duodenum, jejunum, colon and rectum tissue, but up-regulated in the stomach and ileum tissue （P〈0.01）; The mRNA expression level of A QPI 1 was increased 1.7 fold in the jejunum tissue. These results suggested the A QPs gene may play a importment role in the regulation of diarrhea in the stomach and small intestines tissues of piglets during weaning-stress.

关 键 词：仔猪 胃肠道 水通道蛋白基因 克隆 表达 

分 类 号：S852.4[农业科学—基础兽医学]