河川沙塘鳢GH基因及侧翼的克隆与生物信息学分析

doi:10.3969/j.issn.1004-1524.2019.09.09

浙江农业学报 ›› 2019, Vol. 31 ›› Issue (9): 1461-1470.DOI: 10.3969/j.issn.1004-1524.2019.09.09

河川沙塘鳢GH基因及侧翼的克隆与生物信息学分析

刘加林^1,2, 刘士力^2,3,4, 蒋文枰², 程顺², 迟美丽², 郑建波², 贾永义², 赵金良^3,4, 尹绍武^1,*, 顾志敏²

1.南京师范大学生命科学学院,江苏南京 210000;
2.浙江省淡水水产研究所农业农村部淡水渔业健康养殖重点实验室/浙江省淡水水产遗传育种重点实验室,浙江湖州 313001;
3.上海海洋大学农业农村部淡水水产种质资源重点实验室,上海 201306;
4.上海海洋大学上海水产养殖工程技术研究中心,上海 201306

收稿日期:2019-03-07 出版日期:2019-09-25 发布日期:2019-10-11
通讯作者: *尹绍武,E-mail: yinshaowu@163.com
作者简介:刘加林(1994—),男,山东菏泽人,硕士研究生,研究方向为鱼类种质资源与遗传育种。E-mail:1961954694@qq.com
基金资助:
湖州市自然科学资金(2017YZ05); 中国水产科学研究院基本科研业务费(2018HY-XKQ02)

Cloning and bioinformatics analysis of GH gene and its flanking region in Odontobutis potamophila

LIU Jialin^1,2, LIU Shili^2,3,4, JIANG Wenping², CHENG Shun², CHI Meili², ZHENG Jianbo², JIA Yongyi², ZHAO Jinliang^3,4, YIN Shaowu^1,*, GU Zhimin²

1. School of Life Science, Nanjing Normal University, Nanjing 210000, China;
2. Key Laboratory of Healthy Freshwater Aquaculture of Ministry of Agriculture and Rural Development / Key Laboratory of Freshwater Aquatic Animal Genetic and Breeding of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China;
3. Key Laboratory of Freshwater Aquaculture Germplasm Resources, Ministry of Agriculture and Rural Development, Shanghai Ocean University, Shanghai 201306, China;
4. Shanghai Aquaculture Engineering and Technology Research Center, Shanghai Ocean University, Shanghai 201306, China

Received:2019-03-07 Online:2019-09-25 Published:2019-10-11

摘要/Abstract

摘要： 利用长片段PCR、基因组步移法及T-A克隆技术,成功克隆出河川沙塘鳢(Odontobutis potamophila)GH基因全长序列,提交至GenBank获得登录号MH717101。河川沙塘鳢GH基因全序列长5 120 bp,5'端和3'端侧翼序列长度分别为920、682 bp,经预测,上游区域含有MF3、GAGA factor、HNF-3alpha、C/EBP、R2、GATA-1等多种转录调控元件;转录单元长3 518 bp,包含4个内含子和5个外显子,4个内含子大小分别为75、333、2 070和121 bp,5个外显子长度分别为150、131、117、138和373 bp,第3内含子长达2 070 bp,其长度是翘嘴鲌(Culter alburnus)第3内含子的4倍以上;开放阅读框区(open reading frame,ORF)为594 bp,编码由197个氨基酸残基组成的蛋白质多肽。河川沙塘鳢与鲈形目、鲉形目、鲤形目、鲇形目等13种鱼类的氨基酸序列同源性比较表明,其与彼氏冰鰕虎鱼(Leucopsarion petersii)同源性最高,为83.8%,与草鱼(Ctenopharyngodon idella)同源性最低,为49.1%,在180~197氨基酸序列处,河川沙塘鳢表现出高度不保守性,与所列鱼类氨基酸排列均完全不一致。该研究结果为进一步研究河川沙塘鳢GH基因的表达、功能及其转录调控特征奠定了分子基础。

关键词: 河川沙塘鳢, 生长激素, 克隆, 序列分析, 生物信息学

Abstract: The full-length sequence of the Odontobutis potamophila GH gene was successfully cloned by using long segment PCR, genome walking method and T-A cloning technology, which was submitted to GenBank to obtain the login number MH717101. The full-length of the GH gene of Odontobutis potamophila is 5 120 bp, the length of the 5'-end and 3'-end lateral sequence is 920 bp and 682 bp. It is predicted that the upstream region contains MF3, GAGA factor, hnf-3alpha, C/EBP, R2, GATA-1 and other transcriptional regulatory elements, respectively. The transcription unit is 3 518 bp long and contains 4 introns and 5 exons. 4 introns are 75, 333, 2 070 and 121 bp; 5 exons are 150, 131, 117, 138 and 373 bp, respectively. The third intron is 2 070 bp, the length was more than 4 times of the culter third intron. The open reading frame area (ORF) is 594 bp, encoding a protein polypeptide composed of 197 amino acid residues. Comparison of amino acid sequence homology between Odontobutis potamophila and Perciformes, Scorpaeniformes, Cypriniformes, Siluriformes with 13 fishes showed that the homology was the highest to Leucopsarion petersii, at 83.8%, and the homology to Ctenopharyngodon idella was the lowest, at 49.1%, the Odontobutis potamophila to show highly nonconservative at 180-197 amino acid sequence and the amino acid arrangement was completely different with the listed fish. The results of this study laid a molecular foundation for further study on expression, function and transcriptional regulation of GH gene in Odontobutis potamophila.

Key words: Odontobutis potamophila, growth hormone, clone, sequence analysis, bioinformatics

中图分类号:

S917

刘加林, 刘士力, 蒋文枰, 程顺, 迟美丽, 郑建波, 贾永义, 赵金良, 尹绍武, 顾志敏. 河川沙塘鳢GH基因及侧翼的克隆与生物信息学分析[J]. 浙江农业学报, 2019, 31(9): 1461-1470.

LIU Jialin, LIU Shili, JIANG Wenping, CHENG Shun, CHI Meili, ZHENG Jianbo, JIA Yongyi, ZHAO Jinliang, YIN Shaowu, GU Zhimin. Cloning and bioinformatics analysis of GH gene and its flanking region in Odontobutis potamophila[J]. , 2019, 31(9): 1461-1470.

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河川沙塘鳢GH基因及侧翼的克隆与生物信息学分析

Cloning and bioinformatics analysis of GH gene and its flanking region in Odontobutis potamophila

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