文献类型：期刊文献

中文题名：油桐桐酸合成酶基因克隆和植物表达载体构建

英文题名：Cloning and Sequence Analysis of FADX Gene from Vernicia fordii

作者：汪阳东[1] 李元[1] 李鹏[1]

第一作者：汪阳东

机构：[1]中国林业科学研究院亚热带林业研究所

年份：2007

卷号：27

期号：2

起止页码：1-5

中文期刊名：浙江林业科技

外文期刊名：Journal of Zhejiang Forestry Science and Technology

收录：北大核心:【北大核心2004】；

基金：浙江省重大项目"能源植物种质资源与高能植物品种选育及中试"(2005C12003);中国林科院科技发展基金项目"生物柴油植物油桐桐酸合成酶相关基因克隆"(2006)

语种：中文

中文关键词：油桐;FADX基因;克隆;载体构建

外文关键词：Verniciafordii; FADX; cloning; bioinformatic analysis; expressing vector

分类号：S794.3

摘要：α-桐酸合成酶是控制亚油酸向α-桐酸(18:3Δ9cis,11trans,13trans)转化的关键酶。本研究以发育中的油桐种子为材料,利用改良TRIzoL法提取总RNA,并根据GenBank中已经登录的α-桐酸合成酶基因FADX的序列,设计特异性引物,采用RT-PCR方法,克隆得到FADX基因全长cDNA,长度为1221bp。通过生物信息学分析结果表明,该序列5’端和3’端非编码区序列长度分别为13bp和47bp,含有一个开放阅读框(14～1174bp),编码386个氨基酸,含有典型的脂肪酸脱氢酶结构域。将所得基因经BamHI和SacI酶切后插入表达载体质粒pBI121,构建反义表达载体pBI121fadx。
a-eleostearic acid synthetase is the important enzyme for converting linoleic acid into a-eleostearic acid. Total RNA was successfully extracted from growing Vernicia fordii seeds with improved TRIzoL method, cDNA of FADX was cloned by RT-PCR. Bioinformatic analysis showed that the cDNA had length of 1221bp an ORF（ 14 -1 174 bp ） which encoded a polypeptide of 386 amino acids, and contained a typical fatty acid dehydrogenation structural domain. In order to introduce the gene into plant, the cDNA was ligated to the pBI121 in the antisense orientation after digested with the enzyme BamH1 and SacI. The new vector named pBI121fadx was still transformed to leaf of tobacco via Agrobacterium tumefaciens system.