小桐子赤霉素20氧化酶的基因克隆及低温下表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2015.10.023

生物技术通报 ›› 2015, Vol. 31 ›› Issue (10): 140-148.doi: 10.13560/j.cnki.biotech.bull.1985.2015.10.023

小桐子赤霉素20氧化酶的基因克隆及低温下表达分析

王海波^1,2,邹竹荣¹,龚明¹

1.云南师范大学生命科学学院生物能源持续开发利用教育部工程研究中心云南省生物质能与环境生物技术重点实验室, 昆明 650500; 2.曲靖师范学院生物资源与环境科学学院, 曲靖 655011

收稿日期:2015-03-02 出版日期:2015-10-28 发布日期:2015-10-28
作者简介:王海波, 男, 博士研究生, 研究方向：植物逆境生物学; E-mail：bocai0406@163.com
基金资助:
国家自然科学基金项目（31260064, 31460059, 31160169, 31460179）

Gene Cloning and Cold-induced Expression Analysis of the Gene Encoding Gibberellin 20 Oxidase from Jatropha curcas

Wang Haibo^{1, 2}, Zou Zhurong¹, Gong Ming¹

1. School of Life Sciences, Yunnan Normal University, Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Key Laboratory of Biomass Energy and Environmental Biotechnology of Yunnan Province, Kunming 650500; 2. College of Biological Resource and Environmental Science, Qujing Normal University, Qujing 655011

Received:2015-03-02 Published:2015-10-28 Online:2015-10-28

摘要/Abstract

摘要： 赤霉素20氧化酶是植物赤霉素生物合成的限速酶, 决定有生物活性的GA₁与GA₄的合成量。基于先前获得的小桐子低温锻炼转录组数据, 以小桐子幼苗的根为材料, 采用RT-PCR技术克隆到小桐子赤霉素20氧化酶基因JcGA20ox的cDNA序列（GenBank登录号KJ670150.1）。该cDNA全长1 307 bp, 含有完整的开放阅读框（1 131 bp）, 编码376个氨基酸, 分子量为43 kD, 理论等电点为6.7。其推导蛋白属于2-ODD家族, 包含2-酮戊二酸双加氧酶结构域（Fe2OG_OXY）。半定量RT-PCR表达分析显示, JcGA20ox在小桐子各组织中都有表达, 但表达水平具有组织特异性, 在茎中表达量较高, 且受低温诱导表达最显著, 而在叶中表达量相对较低。

关键词: 小桐子, 赤霉素20氧化酶, 2-酮戌二酸双加氧酶结构域, 低温诱导

Abstract: Gibberellin（GA）20 oxidase, a rate-limiting enzyme in GA biosynthesis, determines the amount of active GA forms, GA₁ and GA₄. Based on our previous transcriptome data of chilling-hardened Jatropha curcas, a new gene encoding gibberellin 20 oxidase（named JcGA20ox）was cloned by RT-PCR from the root of J. curcas seedlings（GenBank accession number：KJ670150.1）. The full-length cDNA of JcGA20ox was 1 307 bp, containing an entire open reading frame（ORF）of 1 131 bp. This ORF encoded a polypeptide of 376 amino acids with the molecular weight of 43 kD and the pI value of 6.7. Bioinformatics analysis showed that the JcGA20ox protein contained a Fe²⁺ and 2-oxoglutarate dioxygenase domain（Fe2OG_OXY）, belonging to the 2-ODD family. Semi-quantitative RT-PCR analysis revealed that JcGA20ox expressed differently in different tissues, with abundant and remarkably cold-induced expression in stem, but scarcely in leaves.

Key words: Jatropha curcas, gibberellin 20 oxidase, 2-oxoglutarate dioxygenase domain, cold induction

王海波,邹竹荣,龚明. 小桐子赤霉素20氧化酶的基因克隆及低温下表达分析[J]. 生物技术通报, 2015, 31(10): 140-148.

Wang Haibo, Zou Zhurong, Gong Ming. Gene Cloning and Cold-induced Expression Analysis of the Gene Encoding Gibberellin 20 Oxidase from Jatropha curcas[J]. Biotechnology Bulletin, 2015, 31(10): 140-148.

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小桐子赤霉素20氧化酶的基因克隆及低温下表达分析

Gene Cloning and Cold-induced Expression Analysis of the Gene Encoding Gibberellin 20 Oxidase from Jatropha curcas

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