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[树鼩白细胞介素-2分子特征分析及全长编码序列克隆]

[Analysis of the molecular characteristics and cloning of full-length coding sequence of interleukin-2 in tree shrews].

作者信息

Huang Xiao-Yan, Li Ming-Li, Xu Juan, Gao Yue-Dong, Wang Wen-Guang, Yin An-Guo, Li Xiao-Fei, Sun Xiao-Mei, Xia Xue-Shan, Dai Jie-Jie

机构信息

Institute of Medical Biology, the Chinese Academy of Medical Science, China.

出版信息

Dongwuxue Yanjiu. 2013 Apr;34(2):121-6. doi: 10.3724/SP.J.1141.2013.02121.

Abstract

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 regions of 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.

摘要

虽然树鼩(Tupaia belangeri chinensis)是研究人类疾病机制的优秀动物模型,但很少有研究在疾病模型评估中检测白细胞介素-2(IL-2)这一重要免疫因子。在本研究中,从树鼩脾脏淋巴细胞的RNA中克隆出一段465 bp的全长IL-2 cDNA编码序列,然后用伴刀豆球蛋白A(ConA)培养并刺激这些淋巴细胞。使用Clustal W 2.0对序列和分子特征进行比较分析,确定树鼩与其他哺乳动物之间IL-2整体结构的相似性。树鼩与人类之间IL-2核苷酸序列的同源性为93%,氨基酸同源性为80%。使用MEGA5.0通过邻接法得到的系统发育树结果表明,树鼩、智人和猕猴之间存在密切的遗传关系。三维结构分析表明,树鼩IL-2大部分区域的表面电荷在树鼩和人类之间相似;然而其N-糖基化位点和局部结构不同,这可能会影响抗体结合。这些结果为未来在树鼩中研究IL-2单克隆抗体提供了基础,从而提高它们作为模型的效用。

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