梅花鹿TLR10基因克隆与序列分析

摘要

为系统了解梅花鹿免疫系统,用Trizo法提取梅花鹿肝脏总RNA,采用兼并引物RT-PCR方法克隆了梅花鹿TLR10基因并进行了系统的生物信息学分析.RT-PCR结果显示,dTLR10基因完整ORF区长2439 bp,编码812个氨基酸(GenBank序列号为:HQ260630).同源分析显示,dTLR10基因序列与其他物种TLR10基因高度同源,与同属于偶蹄目反刍类的牛和羊同源性高达95％以上.蛋白功能结构域分析显示,不同物种间TLR10蛋白结构域数量及分布上高度相似,蛋白三维结构预测比较显示,dTLR10蛋白胞外区结构与人类TLR10结构相似,两者均表现为经典的马蹄形结构.%The experiment was conducted to clone Toll-like receptor 10 gene (TLR10 gene) from sika deer and the TLR10 gene was analyzed by bioinformatics software.The total RNA from liver tissue was extracted by Trizol.The TLR10 gene was cloned by RT-PCR with degenerate primer,and TLR10 gene was analyzed by bioinformatics software systematically.The result of RT-PCR showed the complete ORF of dTLR10 gene is 2 439 bp in length,coded 812 amino acids protein (Gen-Bank Number:HQ260630).Homology analysis showed dTLR10 gene is highly homological to others coming from other mammal species,and dTLR10 has more than over 95％ similarity with cattle and sheep belonging to Artiodactyla rumination beasts.The functional domain analysis showed that TLR10 proteins from all species have high similarity in domain number and distribution.Tertiary structure prediction showed the extracellular region of dTLR10 protein is similar to human TLR10 protein with the horseshoe-shape structure.