小桐子甘油-3-磷酸酰基转移酶(JcGPAT)cDNA的克隆与序列分析

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

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1672bp,开放阅读框为1125bp,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。

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.