藏鸡磷酸酪氨酸互作结构域1(PID1)的克隆及组织表达谱研究  

Research on Cloning and Tissue Expression Profile of Phosphotyrosine Interaction Domain Containing 1(PID1)in Tibetan Chicken

作　　者：聂晓庆[1] 林亚秋[1] 徐亚欧[1] 赵燕英[1] 吕明[1] 左璐璐[1] 张小玉[1] 李想[1] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041

出　　处：《畜牧兽医学报》2016年第6期1102-1111,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川应用基础项目(2013JY0044);四川省畜禽育种攻关项目(2011NZ0099-6);西南民族大学项目(2012NFW001)

摘　　要：本研究旨在阐明藏鸡PID1基因在不同组织与时序表达谱,并研究PID1基因表达与肌内脂肪含量(IMF)的关系。利用RT-PCR技术克隆藏鸡PID1基因,用相关软件预测PID1蛋白的结构和功能,采用荧光定量PCR研究PID1基因在藏鸡不同组织、不同日龄的表达谱,并分析其与IMF的相关性。结果显示,克隆得到藏鸡PID1基因序列长度为654bp(GenBank登录号:KT000001),共编码217个氨基酸,是具有PTB结构域的不稳定亲水酸性蛋白质;有14个磷酸化位点、4个O-糖基化位点、1个N-糖基化位点、7种保守特异性蛋白激酶结合位点、3个二硫键;预测其二级结构中α-螺旋占26.73%,β折叠占20.74%,无规则卷曲占52.53%,属于混合型蛋白且主要存在于细胞质中。荧光定量PCR结果显示,PID1基因在藏鸡的不同组织中均存在表达,且在脂肪组织中表达水平最高(P<0.01);时序表达谱结果显示,PID1基因在1日龄公藏鸡胸肌中表达水平最高。在210日龄的公鸡腿肌的表达量极显著高于其他日龄的表达水平(P<0.01),在119日龄母鸡腿肌的表达量极显著高于其他日龄表达水平(P<0.01)。在119和154日龄公鸡脂肪组织中表达水平较高,在210日龄母鸡脂肪组织中的表达量最高(P<0.01)。相关性分析结果显示,PID1mRNA的表达量与藏鸡胸肌的IMF含量存在显著相关(P<0.05)。结果提示,PID1基因可能是影响藏鸡IMF沉积的候选基因。In the present study,PID1 mRNA expression profile in several tissues of Tibetan chicken during different growth phases and the correlation between PID1 mRNA expression and intramuscular fat（IMF）content were investigated.PID1 gene of Tibetan chicken was cloned using RT-PCR and the structure as well as function of PID1 protein was predicted using relevant softwares.Furthermore,the expression profile of PID1 gene was investigated by qRT-PCR and the relationship between PID1 mRNA expression and IMF content in chicken muscles were analyzed.The results showed that PID1 gene was 654 bp encoding a unstable,hydrophilic,acidic protein with 217 amino acids and a PTB（phosphotyrosine binding）domain.There were 14 phosphorylation sites,4 O-glycosylation sites,1 N-glycosylation site,7protein kinase phosphorylationsites,and 3disulfide bonds within the PID1 protein.The predicted secondary structure of PID1 protein was composed of alpha helix（26.73%）,beta fold（20.74%）and random coil（52.53%）,belonging to one of mixed proteins in cytoplasm.qRT-PCR results showed that PID1 mRNA could be expressed in various tissues,with the highest expression level in fat（P〈0.01）.The temporal expression showed that the expression level of PID1 gene was the highest in breast muscle of 1day Tibetan chicken,and was the highest in leg muscle of the 210^th day cocks（P〈0.01）,while the highest in leg muscle of the 119^th day hens（P〈0.01）.For fat tissues,the highest expression level appeared in the 119^th day and the 154^th day cocks,and the 210^th day hens,respectively.The correlation analysis indicated that there was a significant correlation between PID1 mRNA expression and breast muscle IMF content in Tibetan chicken（P〈0.05）.These results suggested that the PID1 gene might play an important role in the IMF deposition of Tibetan chicken.

关 键 词：藏鸡 磷酸酪氨酸互作结构域1 克隆 组织表达 时序表达 肌内脂肪 

分 类 号：S831.2[农业科学—畜牧学]