摘要
利用RT-PCR方法从广西三黄鸡外周血淋巴白细胞中扩增出鸡Toll样受体3基因(TLR3),进而对其进行序列分析和结构预测。结果显示,扩增出的TLR3基因全长为3 036bp,与GenBank中鸡TLR3序列同源性达96.5%～99.7%,突变主要集中在胞外区前173个氨基酸;与牛、鼠、绵羊、猪、马、家兔、黑猩猩、人、大猩猩的氨基酸同源性较低,为68.0%～74.8%;禽类TLR3与其他动物类及人类的TLR3分属于两大不同分支,处于不同进化位置;结构预测表明,GX-sh-chTLR3具有明显的跨膜蛋白结构,胞外区蛋白是由外侧多个α螺旋和内侧凹陷的多个β折叠构成螺线管状马鞍形结构,胞外区含有15个明显的富含亮氨酸的重复序列。结果表明,成功扩增出的鸡TLR3基因编码蛋白的空间结构特征明显,为进一步研究该基因的功能奠定了基础。
In this study,the full-length chicken toll-like receptor 3 gene（TLR3） was amplified from the PBLs of Guangxi Sanhuang chicken by reverse transcription-polymerase chain reaction（RT-PCR）.Sequence analysis and structure prediction of TLR3 gene were carried out.In results,the full-length TLR3 gene was 3 036 bp in size and shared from 96.5% to 99.7% homology with the other TLR3 sequences.The amino acid sequences showed that the mutations mainly happened in 5′ end of extracellular region（from 1 to 173 amino acids）.TLR3 shared a low homology from 68.0% to 74.8% with Bos taurus,Mus musculus,Ovis aries,Suis scrofa,Equus caballus,Oryctolagus,Pan trogldytes,Homo sapiens and Gorilla.In the phylogenetic tree,TLR3s of avian,mammals and rats were divided into two clades.The avian TLR3 belongs to one clade and the mammals and rats TLR3 belong to another clade.Structure prediction showed that TLR3 was a transmembrane protein whose extracellular region showed a horseshoe-shaped solenoid structure with fifteen leucine-rich repeats,consisted of a lot of α-helix on outside and β-sheet on the concave surface inside and they arranged parallelly and alternately.These results suggested the chicken TLR3 gene was distinct in dimentional structure,which might serve as a basis for further study on the function of TLR3 gene.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2011年第6期614-619,共6页
Chinese Veterinary Science
基金
广西青年科学基金项目(桂科青0991014)
广西民族大学2010年研究生教育创新计划项目(gxun-chx2010082)
国家自然科学基金项目(30860206)
关键词
鸡
TLR3
扩增
序列分析
结构预测
chicken
toll-like receptor 3
amplification
sequence analysis
structure prediction
