海南五指山猪TLR2基因克隆及生物信息学分析  

Cloning and bioinformatics analysis of TLR2 gene of Wuzhishan pig from Hainan

作　　者：张艳 黄丽丽 刘海隆 林哲敏[1,2] 谭树义 

机构地区：[1]海南省农业科学院畜牧兽医研究所,海口571100 [2]海南省热带动物繁育与疫病研究重点实验室,海口571100

出　　处：《黑龙江畜牧兽医》2017年第9期37-40,44,286,287,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31560696);海南省自然科学基金项目(20153080);海南省农业科学院农业科技创新专项(CXZX201504)

摘　　要：为了探讨海南五指山猪TLR2基因的分子结构特征及相关遗传变异,为研究TLR2基因遗传变异与免疫疾病的关系打下基础,试验采用PCR技术对该基因组序列进行克隆、测序及相关生物信息学分析。结果表明:获得2 649 bp的五指山猪TLR2基因的DNA序列,其基因编码区全长2 358 bp,编码785个氨基酸;与Gen Bank中发布的普通猪TLR2基因序列相比,共发现9处突变,其中在编码区序列存在2处突变,属于有义突变;与普通猪、马、牛、绵羊、人、黑猩猩、猕猴、犬、小鼠、鸡、斑马鱼的氨基酸序列的一致性分别为99.7%、82.1%、81.4%、80.6%、78.4%、78.4%、77.8%、77.3%、68.2%、50.7%、40.4%;五指山猪TLR2编码的蛋白主要位于质膜,属于分泌性蛋白、亲水性蛋白和跨膜蛋白,有信号肽和5个糖基化修饰位点,存在较多的丝氨酸(Ser)和苏氨酸(Thr)磷酸化位点;不同物种TLR2基因在进化过程中具有较高的保守性。说明TLR2基因可作为今后研究海南五指山猪免疫疾病的重要指标之一。A study was conducted to research the structure and correlated genetic variation of TLR2 gene in Wuzhishan pig,and provided reference for the relation of genetic variation in Hainan Wuzhishan pig TLR2 gene and immunological diseases. PCR and bioinformatics were used to clone,sequence the gene,and analyze in the experiment. The results showed that the 2 649 bp Wuzhishan pig TLR2 DNA sequence was gained,the gene coding region was 2 358 bp( coding 785 amino acid). Compared with the common pig TLR2 DNA sequence,9 mutation sites were found in Wuzhishan pig TLR2 DNA sequence,and manifested 2 mutation sites in CDS,which were sense mutations. The sequence multi-aligned results showed that Wuzhishan pig shared 99.7%,82.1%,81.4%,80.6%,78.4%,78.4%,77.8%,77.3%,68.2%,50.7% and40.4% of similar amino acid sequence with that of Sus scrofa,Equus caballus,Bos taurus,Ovis aries,Homo sapiens,Pan troglodytes,Macaca mulatta,Canis lupus familiaris,Mus musculus,Gallus gallus,Danio rerio,respectively. The protein structure analysis results showed that Wuzhishan pig TLR2 protein attributed secreted,hydrophilic,transmembrane protein,mainly localized plasmalemma. It manifested signal peptide,5 glycosylation sites and more phosphorylation sites in TLR2. The phylogenetic tree of amino acid indicated that TLR2 was highly conserved in the process of evolution of different species. The results indicated that the TLR2 gene can be used as one of indicators in immunological diseases of Hainan Wuzhishan pig.

关 键 词：海南 五指山猪 TLR2基因 克隆 生物信息学分析 序列分析 

分 类 号：S828.89[农业科学—畜牧学] Q78[农业科学—畜牧兽医]