鲤鱼白细胞介素10基因组DNA克隆及序列分析

›› 2013, Vol. 40 ›› Issue (4): 65-68.

鲤鱼白细胞介素10基因组DNA克隆及序列分析

冯祥汝¹, 陈义龙¹, 孙真¹, 贾生美¹, 王文东², 张俊辉¹, 杨振国¹, 卢强¹

1. 吉林大学人兽共患病研究所,吉林长春 130062;
2. 吉林大学畜牧兽医学院,吉林长春 130062

收稿日期:2012-11-28 出版日期:2013-04-20 发布日期:2013-04-19
通讯作者: 卢强(1970-),男,硕士生导师,教授,从事人兽共患病毒病与分子免疫学研究。E-mail:qlu@jlu.edu.cn;Tel:0431-87836717 E-mail:qlu@jlu.edu.cn
作者简介:冯祥汝(1988-),女,辽宁人,硕士生,研究方向:兽医公共卫生学。
基金资助:
国家自然科学基金项目(30972277);吉林大学基本科研业务费项目(200903250)。

Cloning and Sequence Analysis of Interleukin 10 Genomic DNA in Common Carp (Cyprinus carpio L.)

FENG Xiang-ru¹, CHEN Yi-long¹, SUN Zhen¹, JIA Sheng-mei¹, WANG Wen-dong², ZHANG Jun-hui¹, YANG Zhen-guo¹, LU Qiang¹

1. Institute of Zoonoses, Jilin University, Changchun 130062, China;
2. College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China

Received:2012-11-28 Online:2013-04-20 Published:2013-04-19

摘要/Abstract

摘要： 为获得鲤鱼白细胞介素10(interleukin 10,IL-10)全长基因组序列,本研究利用鲤鱼IL-10全长cDNA序列(GenBank登录号:JX524550),通过在两端非编码区设计引物,以提取的鲤鱼脾脏基因组为模板,使用PCR方法成功获得鲤鱼IL-10全长基因组序列。鲤鱼IL-10基因组全长2176 bp,GenBank登录号为JX524551。将所获得的序列与其他物种IL-10基因组序列相比,结果发现都含有5个外显子,4个内含子,外显子在进化上相对保守,剪切位点都符合"gt......ag"规则;序列包含540 bp的开放阅读框,编码179个氨基酸,有2段典型的IL-10氨基酸信号基序。

关键词: 鲤鱼; 白细胞介素10; 基因组; 克隆; 序列分析

Abstract: The purpose of this study was to obtain the full-length sequence of common carp interleukin 10(IL-10) genomic DNA. The sequence of common carp IL-10 full-length cDNA(GenBank:JX 524550) was used. In this study, we designed two pairs of primers in its untranslated regions. Using the carp spleen genomic DNA as the template, we carried out PCR and obtained the IL-10 full-length genomic DNA sequence. The sequence was 2176 bp in length, and the GenBank accession number was JX524551. We compared the gained sequence with the other IL-10 and found their similarity of all harboring 5 exons and 4 introns. Moreover, all exons were comparatively conserved. The exons and introns splice sites were in accordance with "gt......ag" rules. Our results indicated that it was the sizes of introns that were crucial in the process of evolution.

Key words: common carp; interleukin 10; genome; cloning; sequence analysis

中图分类号:

Q785

冯祥汝, 陈义龙, 孙真, 贾生美, 王文东, 张俊辉, 杨振国, 卢强. 鲤鱼白细胞介素10基因组DNA克隆及序列分析[J]. , 2013, 40(4): 65-68.

FENG Xiang-ru, CHEN Yi-long, SUN Zhen, JIA Sheng-mei, WANG Wen-dong, ZHANG Jun-hui, YANG Zhen-guo, LU Qiang. Cloning and Sequence Analysis of Interleukin 10 Genomic DNA in Common Carp (Cyprinus carpio L.)[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(4): 65-68.

参考文献

1 Fiorentino D F, Bond M W, Mosmann T R. Two types of mouse T helper cell. Ⅳ. Th2 clones secrete a factor that inhibits cytokine production by Th1 clones. J Exp Med, 1989, 170:2081~2095.
2 Hedrick J A, Helms A, Vicari A, et al. Characterization of a novel CC chemokine, HCC-4, whose expression is increased by interleukin-10. J Blood, 1998, 91:4242~4247.
3 Inoue Y, Kamota S, Ito K, et al. Molecular cloning and expression analysis of rainbowtrout (Oncorhynchusmykiss) interleukin-10 cDNAs. J Fish Shellfish Immunol, 2005,18:335~344.
4 Moore K W, de Waal M R, Coffman R L, et al. Interleukin-10 and interleukin-10 receptor. J Annu Rev Immunol, 2001, 19:683~765.
5 Kim J M, Brannan C I, Copeland N G, et al. Structure of the mouse IL-10 gene and chromosomal localization of the mouse and human genes. J Immunol, 1992, 148:3618~3623.
6 O’arra A, Vieira P. T(H)1 cells control themselves by producing interleukin-10. J Nat Rev Immunol, 2007, 7:425~428.
7 Pinto R D, Nascimento D S, Reis M I, et al. Molecular characterization, 3D modelling and expression analysis of sea bass (Dicentrar-chus labrax L.) interleukin-10. J Mol Immunol, 2007, 44:2056~2065.
8 Rothwell L, Young J R, Zoorob R, et al. Cloning and characterization of chicken IL-10 and its role in the immune response to Eimeria maxima. J Immunol, 2004, 173:2675~2682.
9 Seppola M, Larsen A N, Steiro K, et al. Characterisation and expression analysis of the interleukin genes, IL-1beta, IL-8 and IL-10, in Atlantic cod (Gadus morhua L.). J Mol Immunol, 2008, 45:887~897.
10 Vieira P, Dang M N, Johnson K E, et al. Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: Homology to Epstein-Barr virus open reading frame BCRFI. J Proc Natl Acad Sci USA, 1991, 88:1172~1176.
11 Zhang D C, Shao Y Q, Huang Y Q, et al. Cloning, characterization and expression analysis of interleukin-10 from the zebrafish (Danio rerion). J Biochem Mol Biol, 2005, 38:571~576.
12 Zou J, Clark M S, Secombes C J. Characterisation, expression and promoter analysis of an interleukin 10 homologue in the puffer fish, Fugu rubripes. J Immunogenetics, 2003, 55:325~335.

[1]	徐扩卫, 冷堂健, 熊宝, 李金燕, 郭盘江, 吴培福, 陈粉粉, 周杰珑. 宁蒗高原鸡全基因组选择信号分析[J]. 中国畜牧兽医, 2025, 52(5): 1941-1954.
[2]	余梦博, 马浩天, 李杨, 潘瑞雪, 龙赟而, 赵一宁, 马驿, 周淑贞. 1株贝莱斯芽孢杆菌G02全基因组测序分析[J]. 中国畜牧兽医, 2025, 52(5): 1977-1986.
[3]	李天秀, 李昕鹏, 董新星, 兰国湘, 严达伟, 朱家位. 丽江猪HMOX2基因扩增、序列分析及组织表达研究[J]. 中国畜牧兽医, 2025, 52(5): 2219-2231.
[4]	刘敏, 程淮, 郑珍珍, 张梦思, 张贺伟, 任静强. 犬副流感病毒N蛋白的真核表达及多克隆抗体制备[J]. 中国畜牧兽医, 2025, 52(5): 2287-2294.
[5]	苗娜, 乔嘉坤, 杨慧, 韩萍萍, 许方军, 车兆轩, 代翔毓, 徐明航, 龙志伟, 朱猛进. 杜洛克猪×二花脸猪F2代杂交猪免疫性状全基因组关联分析[J]. 中国畜牧兽医, 2025, 52(4): 1455-1467.
[6]	刘嗣睿, 刘洪飞, 刘大鹏, 牟启铭, 唐贺贺, 张鹤, 张永福, 侯水生, 周正奎. 整合基因组和转录组筛选调控北京鸭脂肪沉积的候选基因[J]. 中国畜牧兽医, 2025, 52(4): 1468-1477.
[7]	周鑫, 杨铖, 朱桂名, 何黔蓉, 李岩, 樊新忠, 宁超, 王丹. 基于最新参考序列UM_NZW_1.0绘制家兔基因组NUMTs图谱[J]. 中国畜牧兽医, 2025, 52(4): 1660-1670.
[8]	张莹, 陈广振, 刘梅, 李红霞, 俞燕, 贾雪波, 李雯倩, 范建华, 马志禹. 半番鸭NMB基因克隆、生物信息学和组织表达分析[J]. 中国畜牧兽医, 2025, 52(4): 1705-1715.
[9]	杨祝良, 罗锦堂, 张珍, 黎剑能, 李福球, 杨秀荣. 基于“桂芯一号”液相芯片的南丹瑶鸡鸡冠性状全基因组关联分析[J]. 中国畜牧兽医, 2025, 52(4): 1716-1728.
[10]	司维, 谭茜宇, 徐玲芸, 罗廷荣, 李晓宁, 顾金燕. H3N2亚型猪流感病毒HA1蛋白的原核表达及多克隆抗体制备[J]. 中国畜牧兽医, 2025, 52(4): 1739-1749.
[11]	洪嘉俊, 肖金华, 张政, 杨雨婷, 方春, 韩旭. sod基因缺失对单增李斯特菌10403S抗氧化应激作用的影响[J]. 中国畜牧兽医, 2025, 52(4): 1776-1783.
[12]	苗书魁, 米晓云, 魏婕, 魏玉荣, 海力且木·买买提依明. G8型牛轮状病毒VP7基因克隆及生物信息学分析[J]. 中国畜牧兽医, 2025, 52(4): 1784-1795.
[13]	茹梦珂, 李穗湘, 吴雪琴, 严钰璋, 王璐, 程海鹏. 1株鸡源奇异变形杆菌的全基因组测序及生物信息学分析[J]. 中国畜牧兽医, 2025, 52(3): 977-989.
[14]	黄念旎, 黄子珂, 李聪聪, 刘晗, 马诗语, 宋果, 蒋亮, 彭为波, 杨联熙, 黄生强. 九疑山兔FGF10基因克隆、生物信息学及组织表达分析[J]. 中国畜牧兽医, 2025, 52(3): 1001-1010.
[15]	高姣姣, 郑楠, 邵伟, 陈贺, 马宪兰, 赵艳坤. 乳源无乳链球菌的分离鉴定、耐药与毒力特征及基因组分析[J]. 中国畜牧兽医, 2025, 52(3): 1301-1316.