《植物生理学报》 2011, 47(2): 181-188

小桐子甘油-3- 磷酸酰基转移酶(JcGPAT) cDNA 的克隆与序列分析

张楠^1,2, 徐荣华², 刘小烛¹, 刘爱忠^2,*

1 西南林业大学资源学院, 昆明650224; 2 中国科学院西双版纳热带植物园, 昆明650223

通信作者：刘爱忠；E-mail: liuaizhong@xtbg.ac.cn；Tel: 0871-51 404 20

摘　要：

甘油-3-磷酸酰基转移酶是植物生物合成储存油脂过程中的关键酶, 对油料作物种子含油量具有重要的限制作用。本 研究以植物甘油-3-磷酸酰基转移酶同源基因的保守区域序列为基础, 设计简并引物, 结合RACE技术, 从能源植物小桐子 种子中克隆获得 JcGPAT 基因的 cDNA 全长序列(GenBank 登录号 HQ395225)。JcGPAT cDNA 核苷酸序列长度为 1 672 bp, 开放阅读框为1 125 bp, 编码375 个氨基酸。该基因具有明显的GPAT 基因结构域, 其编码的氨基酸序列与油桐、蓖麻等植 物具有很高的同源性。RT-PCR 表达分析表明, 该基因在小桐子发育的种子、叶、根尖等多个组织表达。

关键词：小桐子; GPAT; 简并引物; 基因克隆; 表达分析

收稿：2010-09-03   修定：2010-10-25

资助：国家自然科学基金(30871548)和中国科学院重点方向性项目(KSCX2-YW-G-035-1)。

Cloning and Sequence Analysis of sn-Glycerol-3-Phosphate Acyltransferase Gene (JcGPAT) from Jatropha curcas L.

ZHANG Nan^1,2, XU Rong-Hua², LIU Xiao-Zhu¹, LIU Ai-Zhong^2,*

¹Department of Bioresources, Southwest Forestry University, Kunming 650224, China; ²Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming 650223, China

Corresponding author: LIU Ai-Zhong; E-mail: liuaizhong@xtbg.ac.cn; Tel: 0871-51 404 20

Abstract:

sn-Glycerol-3-phosphate acyltransferase (GPAT), a key enzyme in lipid metabolism, plays a critical role in biosynthesis of lipids in plants. In this study, based on the conserved regions of GPAT genes available from GenBank database, we designed degenerate primers and obtained the cDNA sequences of GPAT gene by RACE technology from Jatropha curcas, named JcGPAT (GenBank accession no. HQ395225). The full length cDNA is 1 672 bp, encoding 375 amino acids which have a high identity (ranging from 78% to 95%) with GPAT genes in other plants reported such as castor (Ricinus communis) and tung tree (Vernicia fordii). RT-PCR analysis showed that JcGPAT was expressed in different tissues including the developing seeds, leaf, root tip and callus.

Key words: Jatropha curcas; GPAT; degenerate primers; gene clone; expression analysis