贵州黑山羊、黔北麻羊IL-2基因的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of IL-2 Gene in Guizhou Black Goat and Qianbei Ma Goat

作　　者：鲜思美[1] 李鹏飞[1] 刘嫒[1] 李婷[1] 包细明 张益[1] 冯将[1] Xian Simei;Li Pengfei;Liu Ai;Li Ting;Bao Ximing;Zhang Yi;Feng Jiang(College of Animal Science of Guizhou University,Guiyang,55002)

机构地区：[1]贵州大学动物科学学院

出　　处：《基因组学与应用生物学》2018年第8期3287-3295,共9页Genomics and Applied Biology

基　　金：贵州省科学技术基金项目[黔科合LH字(2014)7667];贵州省动物疫病防控与兽医公共卫生保障科技创新人才团队[黔科合人才团队(2015)4016号]共同资助

摘　　要：为获得贵州特色品种黑山羊与黔北麻羊IL-2基因,本试验以Con A刺激的贵州黑山羊与黔北麻羊外周血淋巴细胞为材料,采用RT-PCR方法分别扩增贵州黑山羊与黔北麻羊IL-2基因,进行克隆与生物信息学分析。结果显示,（1）贵州黑山羊与黔北麻羊IL-2基因核苷酸与氨基酸一致性为100%,完整编码阅读框大小为468 bp,编码155个氨基酸;（2）黔北麻羊、贵州黑山羊IL-2基因与Gen Bank中羊源IL-2基因核苷酸一致性为95.9%~100%,氨基酸一致性为91.1%~100%;（3）遗传进化树结果表明,贵州黑山羊与黔北麻羊IL-2基因处于同一进化分支,与羊源IL-2基因亲缘关系较近;（4）生物信息学分析结果显示,IL-2蛋白二级结构中以α螺旋与无规则卷曲较多,该蛋白为亲水性蛋白,含信号肽序列,无跨膜结构,无特征功能结构域;（5）不同动物IL-2蛋白的抗原性分析存在差异,贵州黑山羊/黔北麻羊IL-2蛋白与羊源IL-2蛋白的比较差异较小,与人和其它动物的比较差异较大。由此得出结论,获得贵州黑山羊与黔北麻羊IL-2基因完整编码区序列,二者核苷酸与氨基酸序列一致。贵州黑山羊/黔北麻羊IL-2基因与羊源IL-2基因亲缘关系近,预测其编码的蛋白为亲水性的分泌型蛋白,不同动物IL-2基因抗原性存在差异。To obtain the IL-2 gene of Black goat and Qianbei ma goat which were two native breed in Guizhou province,this research took the peripheral blood mononuclear cells as a material,which was from Black goat and Qianbei ma goat and stimulated by Con A.And we amplified the IL-2 gene respectively by RT-PCR,then cloning and bioinformatics analysis were carried out.The results showed that the IL-2 gene of Guizhou Black goat and Qianbei ma goat were the same at the nucleotide sequence and amino acid sequence,and its ORF was 468 bp,which encoded 155 amino acids.The nucleotide sequences similarity of two IL-2 genes with the IL-2 gene in Gen Bank were 95.9%~100%,and the amino sequences similarity were 91.1% ~100%.The phylogenetic tree showed that the IL-2 genes in Guizhou Black goat and Qianbei ma goat were from the same evolutionary branch,which were close with the IL-2 gene in Gen Bank.The bioinformatics analyses showed that theα-helix and random coil were major in the IL-2 protein secondary structure.And the protein was hydrophilic with signal peptide sequence,but no transmembrane structure and functional structure domain.Comparative analysis of antigenicity showed that there had difference of the IL-2 protein among different species.The IL-2 protein difference between Guizhou Black goat/Qianbei ma goat and other sheep derived were small,but the difference between Guizhou black goat/Qianbei ma goat and people or other animals was great.The IL-2 coding sequence of Guizhou Black goat and Qianbei ma goat were acquired,indicating the nucleotide and amino acid sequences were similar.They might have a close relationship with the IL-2 gene of sheep derived animals.The coding protein of IL-2 might be the secreting type and possess hydrophily,and the antigenicity of different animals existed difference.

关 键 词：贵州黑山羊 黔北麻羊 IL-2基因 克隆与分析 

分 类 号：S827[农业科学—畜牧学]