牦牛Toll样受体8和9基因克隆及组织表达分析  

Cloning and Expression Analysis of Yak Toll-like Receptor 8 and 9 Genes

作　　者：黄勇[1] 兰道亮[2] 陈亚冰[1] 林宝山[1] 黄偲[1] 李键[2] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]西南民族大学青藏高原研究院,四川成都610041

出　　处：《动物医学进展》2015年第6期34-38,共5页Progress In Veterinary Medicine

基　　金：中央高校青年教师基金项目(2014NZYQN37);西南民族大学研究生创新型科研项目(CX2015SZ065)

摘　　要：为了解牦牛Toll样受体8(TLR8)和9(TLR9)蛋白的结构特征及其基因的组织表达规律,丰富牦牛TLRs基因研究的理论数据,应用RT-PCR方法克隆牦牛TLR8、TLR9基因,并进行生物信息学分析,采用实时荧光定量PCR技术构建组织表达谱。结果显示,两个基因cDNA全长分别为3 102bp和3 090bp。牦牛TLR8和TLR9基因与野牦牛、黄牛和绵羊之间的同源性都很高,均在95%以上。牦牛TLR8和TLR9蛋白都具有胞外LRRs功能域、胞内区TIR结构域、低复杂度区;另外,TLR8还具有1个C端富集亮氨酸重复序列(LRRCT)和跨膜结构域。两个基因在10个组织中均有不同程度的表达,在肾脏中表达量最高,而在大肠中表达量最低。本研究成功克隆了牦牛TLR8和TLR9基因的完整编码区,并揭示了它们在各组织器官的表达规律,为进一步研究TLRs在牦牛体内的免疫机制奠定了基础。In order to understand the structural characteristics of yak Toll-like receptor 8 and 9 proteins and their gene expression patterns, as well as enrich the theory research of TLRs genes in yak, this study used RT-PCR method to clone TLR8 and TLR9 genes of yak. Moreover, the bioinformatics software was used to analyze the characteristics and real-time PCR technique was used to construct the gene expression spectram of TLR8 and TLR9. The results showed that TLR 8 and 9 have coding sequences of 3 102 bp and 3 090 bp which encode a pro- tein of 1033 and 1029 amino acids, respectively. The nucleotide sequence homology analysis showed the genes in yak have a very high similarity with Bos mutus, cattle and sheep. Both TLR8 and TLR9 proteins of yak were pre- dicted to have extracellular leucine-rich repeats domain, cytosolie TIR （Toll/IL-1 receptor homologous region） do- main and one low complexity region. In addition, TLR8 has a C-terminal of leucine-rich repeats domain （LRRCT） and transmembrane domains. These two genes have different levels of expression in ten tissues examined, either of them have highest level of expression in kidney and lowest in large intestine. This study successfully cloned the complete coding region of TLR8 gene and TLR9 gene in yak and examined the expression patterns of TLRs in vari- ous tissues and organs, which provide theoretical foundation for further study on TLRs in the immune mechanism of yak.

关 键 词：牦牛 Toll样受体8 TOLL样受体9 克隆 表达 

分 类 号：Q78[生物学—分子生物学] S858.23[农业科学—临床兽医学]