牦牛TLR6基因的克隆、序列分析及其表达分布研究  

Cloning, sequence analysis of TLR6 gene of yak and expression and distribution in tissues

作　　者：林宝山[1] 兰道亮[2] 黄偲[1] 陈亚冰[1] 张雁[1] 王英 李键[1,2] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041 [2]西南民族大学青藏高原研究院,成都610041 [3]四川省动物疾病预防控制中心,成都610041

出　　处：《东北农业大学学报》2014年第7期91-97,103,共8页Journal of Northeast Agricultural University

基　　金：中央高校青年教师基金项目(2014NZYQN37)

摘　　要：为了解牦牛TLR6基因特点及其在牦牛各组织动态表达分布规律,根据GeneBank中野牦牛TLR6序列设计引物克隆并对所得序列进行生物信息学分析,建立荧光定量PCR方法,对TLR6基因在牦牛组织中的表达分布进行研究。基因序列分析表明,克隆得到牦牛TLR6基因编码区全长2 397 bp,编码氨基酸798个,N端含有26个氨基酸组成的信号肽,分子质量91.5719 ku,理论等电点6.45。蛋白预测结果表明,TLR6编码蛋白整体表现为疏水性。同源性分析表明,10条序列当中牦牛与野牦牛、普通牛、藏羚羊等物种具有较高同源性,在系统发育树中距离最近。该结果说明TLR6基因序列具有较高保守性。荧光定量结果表明,TLR6基因在牦牛所有组织中均有表达,TLR6基因在脾脏表达量最高,在肌肉表达量最低,在卵巢、肺、小肠、肝、肾、心等组织依次有较高表达。该结果可为进一步研究TLRs在牦牛等其他高原动物体内的分布及动态表达规律,以及牦牛抗病免疫及育种奠定基础。To understand the characteristics of yak TLR6 gene and the dynamic distribution in various tissues and organs. According to Bos mutus TLR6 sequence in GeneBank, The paper designed the cloning and real-time PCR primers. Besides, we successful y established a real-time PCR methods for detecting the expression pattern of TLR6 gene in yak. The results showed that the length of the cloned cDNA of TLR6 was 2 397 bp, it encoded a peptide of 798 amino acids whose calculated molecular weight was 91.5719 ku, PI was 6.45. The protein prediction software showed that TLR6 protein performed hydrophobicity. Sequence homology analysis showed that yak had higher homology with Bos mutus, Bos taurus, Pantholops hodgsoni and other species among the nine sequences, which had the nearest perimeter in the phylogenetic tree. The results of real-time PCR showed that TLR6 gene was expressed in al the tissues of yak, the highest in the spleen, the lowest in the muscle and fol owed by the lung, ovary, smal intestine and other tissues. The results suggested that TLR6 gene might play an important role in the immune molecular mechanisms of yak disease.To study the distribution and dynamics expression of TLRs in yaks and other animals in the highland is of significance, and provides a basis for future research on the breeding for disease resistance of yak.

关 键 词：牦牛 TLR6 克隆 荧光定量PCR 组织表达分布 

分 类 号：S823.85[农业科学—畜牧学] Q785[农业科学—畜牧兽医]