羊IL-1Ra基因全长cDNA克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.03.005

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (3): 598-603.doi: 10.16431/j.cnki.1671-7236.2018.03.005

羊IL-1Ra基因全长cDNA克隆及生物信息学分析

水一鸣¹, 刘楠楠¹, 郭兴², 邹德颖², 翟菲菲¹, 王路鹿¹, 鞠丹迪¹, 张士军¹, 胡盼¹, 李岩松¹, 卢士英¹, 周玉¹, 柳增善¹, 李兆辉¹, 任洪林¹

1. 吉林大学动物医学学院, 吉林大学人兽共患病研究所, 人兽共患病研究教育部重点实验室, 长春 130062;
2. 盘锦检验检测中心, 盘锦 124010

收稿日期:2017-09-14 出版日期:2018-03-20 发布日期:2018-03-22
通讯作者: 李兆辉, 任洪林 E-mail:cclzh@126.com;renhl@jlu.edu.cn
作者简介:水一鸣(1992-),女,安徽马鞍山人,硕士生,研究方向:人兽共患病,E-mail:seechul@163.com
基金资助:
吉林省科技发展计划（20150204078NY、20170101149JC）

Cloning and Bioinformatics Analysis of IL-1Ra Gene Full-length cDNA in Sheep

SHUI Yiming¹, LIU Nannan¹, GUO Xing², ZOU Deying², ZHAI Feifei¹, WANG Lulu¹, JU Dandi¹, ZHANG Shijun¹, HU Pan¹, LI Yansong¹, LU Shiying¹, ZHOU Yu¹, LIU Zengshan¹, LI Zhaohui¹, REN Honglin¹

1. Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China;
2. Panjin Inspection and Testing Center, Panjin 124010, China

Received:2017-09-14 Online:2018-03-20 Published:2018-03-22

摘要/Abstract

摘要：

试验旨在对羊白介素1受体颉颃因子（interleukin-1 receptor antagonist，IL-1Ra）基因进行全长cDNA克隆及生物学分析。根据布鲁氏菌感染羊白细胞层SSH cDNA文库中的IL-1Ra基因序列信息及已知核苷酸序列（GenBank登录号：KC425613.1）设计引物，利用RT-PCR结合RACE方法扩增并克隆IL-1Ra基因，对其进行序列及相关分子特性分析。结果表明，羊IL-1Ra基因全长为1 228 bp，可编码174个氨基酸，含有1个完整的IL-1保守结构域和1个IL-1Ra结构域（PHA02651结构域）。IL-1Ra分子质量为19 765.8 u，等电点（pI）为5.72，其分子式为C₈₈₅H₁₃₈₅N₂₃₅O₂₅₆S₁₁，且含有信号肽。二级结构分析显示，IL-1Ra分子存在较多的β折叠及受体结合位点，与三级结构预测结果一致，且与人/鼠IL-1Ra分子的空间结构相似度极高，达到90%以上。羊IL-1Ra有IL-1特有的三叶草结构，其酶结合结构域位于TYR⁴⁷至GLU⁶⁶之间。将羊IL-1Ra与牛、虎鲸、宽吻海豚、猪、家犬、人、鼠等14个物种进行蛋白质同源性比对，发现在各物种间存在5个高度同源的半胱氨酸位点。系统进化树分析发现，羊IL-1Ra基因全长cDNA所编码的氨基酸序列同山羊、牛单独形成一个分支。本研究结果为今后深入研究IL-1Ra基因的生物学功能奠定了基础。

关键词: 羊; IL-1Ra基因; 生物信息学; 分析

Abstract:

This study was aimed to clone ovine interleukin-1 receptor antagonist (IL-1Ra) gene and analyze the gene structure using bioinformatics.The primers was designed according to the obtained SSH cDNA library IL-1Ra gene sequence information and the known nucleotide sequence(GenBank accession No.:KC425613.1),IL-1Ra gene was amplified and cloned by RT-PCR and RACE methods,IL-1Ra sequence and its related molecular characteristics were analyzed.The results showed that the length of IL-1Ra gene was 1 228 bp,and encoded 174 amino acids.It was predicted that IL-1Ra had a complete IL-1 conserved domain and IL-1Ra domain (PHA02651 domain).The molecular weight of IL-1Ra was 19 765.8 u,the isoelectric point (pI) was 5.72,its molecular was C₈₈₅H₁₃₈₅N₂₃₅O₂₅₆S₁₁.IL-1Ra contained the signal peptide.The secondary structure of IL-1Ra had more β-fold and receptor binding sites,which were consistent with the predicted results of tertiary structure,and had a similarity to the spatial structure of human/mouse IL-1Ra,up to more than 90%.The IL-1Ra contained a clover structure which was peculiar to the IL-1,and its enzyme-binding domain was located between TYR⁴⁷ and GLU⁶⁶.The results showed that there were five highly homologous cysteines between sheep and the other fourteen species.By establishing the phylogenetic tree of IL-1Ra,it was found that the full-length cDNA of IL-1Ra in sheep formed a branch with goat and cattle.This study result could provide the basis for the further study of the biological function of IL-1Ra gene.

Key words: sheep; IL-1Ra gene; bioinformatics; analysis

中图分类号:

Q785

水一鸣, 刘楠楠, 郭兴, 邹德颖, 翟菲菲, 王路鹿, 鞠丹迪, 张士军, 胡盼, 李岩松, 卢士英, 周玉, 柳增善, 李兆辉, 任洪林. 羊IL-1Ra基因全长cDNA克隆及生物信息学分析[J]. 《中国畜牧兽医》, 2018, 45(3): 598-603.

SHUI Yiming, LIU Nannan, GUO Xing, ZOU Deying, ZHAI Feifei, WANG Lulu, JU Dandi, ZHANG Shijun, HU Pan, LI Yansong, LU Shiying, ZHOU Yu, LIU Zengshan, LI Zhaohui, REN Honglin. Cloning and Bioinformatics Analysis of IL-1Ra Gene Full-length cDNA in Sheep[J]. , 2018, 45(3): 598-603.

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羊IL-1Ra基因全长cDNA克隆及生物信息学分析

Cloning and Bioinformatics Analysis of IL-1Ra Gene Full-length cDNA in Sheep

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