莱芜猪PID1基因的功能分析及表达谱研究

摘要

In order to understand the structure and function of porcine PID1 gene, the molecular structure and function of porcine PID1 from the Laiwu pig sample were analyzed. The mRNA expression profile of the porcine PID1 gene in ten tissues (heart, longissimus dorsi, spleen, liver, lung, small and large intestine, brain, fat and spinal) were examined by QRT-PCR. Bioinformatic analysis revealed that the porcine PIDI protein was located in the cytoplasmic, encoded 217 amino acids with a molecular weight of 54 ku and a pl of 5. 11. PID1 was a non-transmembrane protein and was an unsecreted and hydrophilic protein, α helix occupied the higher proportion in secondary structure. In addition, PID1 gene had a phosphotyrosine-binding (PTB) domain on the C terminal of protein. Laiwu pig, homo spiens and pan troglodytes were closed in the phylogenetic tree. Real-time PCR analysis showed that the PID1 gene was expressed in various tissues, but PID1 mRNA abundances, on the whole, tended to be heart ＞ brain ＞ liver ＞ fat ＞longissimus dorsi ＞ lung＞ spinal＞ large intestine＞ spleen, small intestine was not detected.The results implied that PID1 may be related to insulin signal pathway and regulate the expression of GLUT4, which showed us its extensive functions for transcription.%为进一步了解莱芜猪PID1基因的结构和功能,本研究以莱芜猪为试验对象,分析PID1基因的结构和功能,并利用SYBR-Green实时荧光定量PCR方法,分析该基因在莱芜猪10个不同组织(心脏、背最长肌、脾脏、肝脏、肺脏、小肠、大肠、脑、脂肪、脊髓)的表达谱信息.生物信息学分析表明:PID1基因编码含217个氨基酸的蛋白,其定位在细胞质,无跨膜结构,为非分泌蛋白,具有亲水性,分子量为54 ku,等电点为5.11,二级结构中α螺旋占很大的比例,PID1的C端有一PTB(磷酸酪氨酸作用位点)结构域,与人和黑猩猩的亲缘关系较近.组织表达谱分析结果显示该基因在大多数组织中均有表达,其在各组织中的mRNA表达丰度表现为:心脏＞脑＞肝脏＞脂肪＞背最长肌＞肺脏＞脊髓＞大肠＞脾脏,而在小肠中未见明显表达.结果提示:PID1可能与胰岛素信号途径有关,并可能调控GLUT4基因的表达,呈现多组织表达特征,提示其调节目的基因转录具有广泛性.