摘要
用RT-PCR技术从日本大耳白兔脾脏组织克隆出兔Toll样受体2、3、4基因(拟命名为RTLR2、RTLR3和RTLR4)的cDNA序列并进行测序,获得的3个Toll样受体基因序列长分别为128、150和139 bp,并将其测序结果与GenBank中登录的穴兔(Oryctolagus cuniculus)的Tolls核苷酸序列进行比对,发现本次克隆到的_RTLR2与穴兔TLR2的基因序列(NM_001082781)相似性为99%,而RTLR3与TLR3(NM_001082219)和RTLR4与TLR4(NM_001082732)相似性均为100%。Protein Blast同源性结果显示,_RTLR2、_RTLR3和_RTLR4的氨基酸序列与穴兔TLR2、TLR3和TLR4的同源性皆为100%,与马、人、野猪等其他8种动物TLRs的比较,与RTLR2同源性最高的是马的80%,其他的只有61%～78%;与RTLR3同源性最高是马和野猪的97%,其他也较高达93%～95%;而RTLR4与其他8种动物的同源性均较低75%以下。结果表明:RTLR2、RTLR3和RTLR4分别为兔TLR2、TLR3和TLR4的部分cDNA基因序列;TLRs在不同物种的进化过程中存在种属特异性。
In this study, Toll-like receptor 2 （R TLR2 ）, Toll-like receptor-3 （R TLR3） and Toll-like receptor-4 （R TLR4 ） gene were cloned from Japanese white rabbits spleen by reverse transcription-polymerase chain reaetion（RT-PCR）. Sequence analysis indicated that the RTLR2,RTLR3 and RTLR4 eDNA cloned were 128,150 and 139 bp in length and that the nucleotide sequence of RTLR2 shared 99 % homology with the published sequence Oryctolagus cuniculus TLR2（NM_001082781）,while RTLR3 and RTLR4 shared 100% homology with the published sequence TLR3 （NM_001082219）and TLR4 （NM_001082732）. The predicted amino acid sequence of RTLR2,RTLR3 and RTLR4 gene was compared to that of the partial cDNA fragments of Oryctolagus cuniculus TLR2, TLR3 and TLR4, with 100% similarity. The comparison of the deduced amino acids sequence of R TLR2,RTLR3 and R TLR4 with that of horse,dog,cat, orangutan,human, cattle sheep and mouse showed that the amino acids homology similarity were 80%,78%,78%,78%,78%,76%,73% and 61% in TLR2,97%,97%,95%,95%,95%,95%,93%and 93% in TLR3, and 75 %, 75 %, 75 %, 73 %,71 %, 66 %,66 % and 62 % in TLR4, respectively. Based on the phylogenetic tree and alignment of predicted animo sequence,we concluded that R TLR2,RTLR3 and RTLR4 were partial eDNA fragments of Oryctolagus cuniculus TLR2,TLR3 and TLR4,individually. Likewise,it was suggested that there was species-specific of TLRs in different kinds of animials.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2012年第12期1804-1807,共4页
Chinese Journal of Veterinary Science
