摘要Toll样受体（TLRs）是一类参与天然免疫的模式识别受体（PRRs），是一类I型跨膜糖蛋白，同时也是连接特异性免疫与非特异性免疫的桥梁。TLR5作为Toll样受体家族成员之一，是机体免疫系统特异性识别细菌鞭毛蛋白的主要受体之一。本实验依据GenBank中已公布的犬类TLR5基因的保守序列设计出特异性引物。PCR扩增获得犬类TLR5目的基因，并构建重组质粒pCR2.1-DogTLR5，测序后对比发现序列正确。实验结果显示，犬类TLR5基因的大小为3130bp，含一个2577bp大小的ORF，可以编码858个氨基酸。核苷酸序列同源性比对结果表明，犬类TLR5基因序列与大熊猫、雪豹、东北虎和猎豹 TLR5 基因同源性相对于其他比对物种较高，分别为78.7%、77.2%、76.8%和76.3%。目前虽然已有一些关于家禽（如猪、鹅、小鼠等）TLR5基因的研究报告，但关于犬类TLR5基因的研究甚少。对于TLR5基因的特征以及序列的研究，为进一步了解犬类非特异性免疫的特点，为进一步在分子水平研究犬类TLR5的结构、分布与功能奠定基础，对更深入阐明犬类TLR5在介导机体免疫反应信号传导机制中的作用具有重要意义。42689
关键词：犬类 TLR5基因 克隆 序列分析
Molecular Cloning and analysis of canine TLR5 gene
Abstract
Toll like receptors are a kind of pattern recognition receptors which involved in nonspecific immunity. They are also a type of I transmembrane glycoprotein.At the same time,TLRs are also a bridge which links nonspecific immunity and specific immunity.TLR5 as a member of TLRs family, is one of the main receptor of the immune system to identify bacterial flagellin.Based on the conservative sequence of TLR5 gene in GenBank, the experimental results of this study were designed with specific primers.PCR amplification to obtain the dog TLR5 target gene, and construct the recombinant plasmid pCR2.1-DogTLR5, after sequencing, the sequence is correct.The experimental results show that the size of TLR5 gene is 3130bp, which contains an open reading frame of 2577bp, which can encode 858 amino acids.Nucleotide sequence homology comparison results show that, the dogs TLR5 gene sequence and the giant panda, snow leopard, Siberian tiger and leopard TLR5 gene homologous relative to other than species is higher, 78.7% and 77.2% and 76.8% and 76.3%, respectively.Although there have been some of research reports about the poultry TLR5 gene(such as pigs, geese and mouse), but the study on canine TLR5 gene is very little.Studies on the characteristics of TLR5 gene and its series, in order to further understand the dog’s non specific immunological characteristics, and lay the foundation for further study of canine TLR5 structure, distribution and the function in molecular level. Meanwhile, it will have great significance to further elucidate the role of canine TLR5 in mediating immune response signal conduction mechanism .
Key word: canine TLR5 clone sequence analysis
目 录
摘要.Ⅰ
Abstract.II
目录.Ⅲ
图清单Ⅳ
表清单Ⅳ
1 绪论..5
2 材料与方法..7
2.1 材料..7
2.2 方法..7
3 结果10
3.1 PCR产物电泳结果.10
3.2 菌液PCR鉴定阳性克隆结果.10
3.3 犬类TLR5基因测序结果.11
3.4 犬类TLR5基因同源性分析12
4 讨论13
5 结论14
参考文献..15
致谢.17
图清单
图序号 图名称 页码
图1-1 TLR家族胞外域结构-Toll样受体与配体复合物结构的研究进展 6 犬类TLR5基因的克隆及分析:http://www.youerw.com/shengwu/lunwen_43200.html