水貂部分Toll样受体(TLRs)基因的分子克隆、序列分析及组织表达分析

Molecular Cloning,Sequence Analysis and Tissues Expression Profile of Part of Mink Toll-like Receptors(TLRs) Genes

试验旨在研究Toll样受体(Toll-like receptors,TLRs)在水貂抗病毒免疫中的作用机制,并为抗病育种积累可供选择的基因素材。本试验以雪貂TLRs基因为参考序列设计4对引物对水貂TLR4、TLR6、TLR7及TLR8进行分子克隆,并对所得序列进行生物信息学分析,进一步利用半定量RT-PCR技术分析TLR4和TLR7基因mRNA在不同年龄水貂各组织中的表达情况。结果表明,水貂TLRs与食肉目动物(雪貂、北极熊、大熊猫、海象、海豹、犬、老虎和猫)具有较高的同源性,在系统发育树中距离最近;组织表达分析表明TLR4和TLR7在检测的组织中广泛表达且表达量存在差异,相比成年水貂,仔貂各组织中TLR4或TLR7基因表达量更高。

The present study was aimed to explore the potential role of Toll-like receptors （TLRs） on the mink immunity, and accumulate alternative genetic material for the mink breeding for dis- ease-resistant. Four pairs of primers were designed according to ferret TLR sequences in GenBank to obtain the sequences of TLR genes （TLR4, TLR6,TLR7 and TLR8） of mink and then per- formed bioinformatics analysis of these sequences. In addition,the expression of TLR4 and TLR7 genes in various tissues in young and adult minks were analyzed by RT-PCR. Sequence homology analysis showed that mink had higher homology with carnivora （ferrets, polar bear, panda,wal- rus, seals,dog,tiger and cat） which had the nearest perimeter in the phylogenetic tree. Tissue ex- pression analysis showed that TLR4 and TLR7 were widely and differently expressed in variety tissues of mink. Interestingly,the expression of TLR4 and TLR7 in young mink were much higher than that in adult mink.