树鼩IL-2全长编码序列的克隆及分子特征分析  

Analysis of the molecular characteristics and cloning of full-length coding sequence of Interleukin-2 in tree shrews

作　　者：黄晓燕[1] 李明利[2] 徐娟[1] 高跃东[3,4] 王文广[1] 殷安国[1] 李晓飞[1] 孙晓梅[1] 夏雪山[5] 代解杰[1] 

机构地区：[1]中国医学科学院/北京协和医学院医学生物学研究所树鼩种质资源中心云南省重大传染病疫苗研发重点实验室,云南昆明650118 [2]中国科学院昆明动物研究所动物模型与人类疾病机理重点实验室:免疫生物学实验室,云南昆明650223 [3]中国科学院昆明生物多样性大型仪器区域中心,云南昆明650223 [4]中国科学院昆明动物研究所遗传资源与进化国家重点实验室,云南昆明650223 [5]昆明理工大学生命科学与技术学院,云南昆明650500

出　　处：《Zoological Research》2013年第2期121-126,共6页动物学研究（英文）

基　　金：国家科技支撑计划项目(2009BAI83B02-21;2011BAI15B01-21;2012BAI39B01);云南省科技计划重点项目(2006PT07-2);云南省应用基础面上项目(2011FZ211)

摘　　要：树鼩作为多种人类疾病模型已受到广泛关注,而免疫因子对于树鼩模型评价至关重要,但目前对其白细胞介素-2(IL-2)的研究鲜有报道。该实验以经ConA(concanavalin)诱导培养的树鼩淋巴细胞总RNA为模板,RT-PCR克隆出465bp的树鼩IL-2全长编码序列,并采用ClustalW软件分析其序列和分子特征。结果表明树鼩IL-2cDNA编码一个由154个氨基酸组成的蛋白质,其cDNA及氨基酸序列与人的同源性分别为93%及80%,且其整体结构与人IL-2相似。MEGA5.0软件构建的进化树表明,树鼩与人及恒河猴的亲缘关系较近。Pymol软件对树鼩和人IL-2氨基酸序列进行的三维结构模建表明,两者的IL-2分子三维空间结构基本相似,表面大部分区域所带电荷相同,但在某些区域差异较大,且树鼩多出一个糖基化位点,这些差异对抗体的结合可能存在影响。该研究为今后树鼩IL-2单克隆抗体的制备及功能研究奠定了基础。While the tree shrew （Tupaia belangeri chinensis） is an excellent animal model for studying the mechanisms of human diseases, but few studies examine interleukin-2 （IL-2）, an important immune factor in disease model evaluation. In this study, a 465 bp of the full-length IL-2 cDNA encoding sequence was cloned from the RNA of tree shrew spleen lymphocytes, which were then cultivated and stimulated with ConA （concanavalin）. Clustal W 2.0 was used to compare and analyze the sequence and molecular characteristics, and establish the similarity of the overall structure of IL-2 between tree shrews and other mammals. The homology of the IL-2 nucleotide sequence between tree shrews and humans was 93%, and the amino acid homology was 80%. The phylogenetic tree results, derived through the Neighbour-Joining method using MEGA5.0, indicated a close genetic relationship between tree shrews, Homo sapiens, and Macaca mulatta. The three-dimensional structure analysis showed that the surface charges in most regionsof tree shrew IL-2 were similar to between tree shrews and humans; however, the N-glycosylation sites and local structures were different, which may affect antibody binding. These results provide a fundamental basis for the future study of IL-2 monoclonal antibody in tree shrews, thereby improving their utility as a model.

关 键 词：树鼩 IL-2 克隆 结构 功能 

分 类 号：R-332[医药卫生]