栽培种花生FAE1基因的克隆和序列分析
2018-08-14张欣孙全喜吴琪王秀贞胡东青唐月异王志伟宋国生徐建志殷冬梅王传堂
张欣 孙全喜 吴琪 王秀贞 胡东青 唐月异 王志伟 宋国生 徐建志 殷冬梅 王传堂
摘要:脂肪酸链延长酶1(FAE1)是控制脂肪酸延长的关键酶,对芥酸的生物合成有至关重要的作用。本研究从栽培种花生的两个染色体组中分别克隆得到2个FAE1基因,命名为FAE1-A和FAE1-B。FAE1只有一个开放阅读框,无内含子,完整编码区长1 536 bp,编码511个氨基酸。利用相关软件或在线工具对FAE1进行生物信息学分析,结果表明其编码蛋白二级结构以α螺旋为主;无信号肽,表明FAE1蛋白不是分泌蛋白,而是在细胞中发挥作用,且主要存在于内质网上;FAE1蛋白不稳定系数小于40,总平均亲水系数为负数,属于不稳定的亲水性蛋白。本研究可为花生低芥酸育种提供依据。
关键词:FAE1;芥酸;花生;基因克隆;生物信息学分析
中图分类号:S565.2:Q781文献标识号:A文章编号:1001-4942(2018)06-0035-06
Abstract Fatty acid elongation 1 (FAE1) is a key enzyme that controls the elongation of fatty acids and plays a crucial role in the biosynthesis of erucic acid. In this study, two FAE1 genes, FAE1-A and FAE1-B, were cloned from two genomes of the cultivated peanut, respectively. FAE1 had only one open reading frame, no introns, and the complete coding sequence of the gene is 1 536 bp, encoding 511 amino acids. The bioinformatics analysis results showed that the secondary structure of FAE1 protein was mainly α-helix; there was no signal peptide, indicating that FAE1 was not secretory protein, and it worked in cells and mainly existed in endoplasmic reticulum; the instability coefficient of FAE1 was lower than 40 and the total mean hydrophilic coefficient was negative, so FAE1 was unstable hydrophilic protein. The study may facilitate peanut breeding for low erucic acid content.
Keywords FAE1; Erucic acid;Peanut; Gene cloning; Bioinformatics analysis
芥酸(C22∶1)是一种常见于十字花科植物种子中的长链脂肪酸,不易被人体消化吸收。试验证明,芥酸积累会引起心肌脂肪沉淀并导致心肌脂肪代谢障碍,严重危害健康[1-3]。因此,低芥酸含量已成为食用油品质的一项重要指标。
FAE1是控制脂肪酸碳链延长的关键基因[4]。该基因所编码的酶是长链脂肪酸合成过程的限速酶,决定了脂肪酸碳链的长度[5]。现已从拟南芥、甘蓝型和芥菜型油菜等植物中克隆到该基因[6-8],虽然不同材料中FAE1基因在长度与序列上存在差异,但均无内含子[9]。目前已将拟南芥FAE1基因转化到酵母、拟南芥、烟草和油菜中,FAE1的表达能使酵母和烟草的长链脂肪酸积累,且擬南芥和油菜基因组中FAE1拷贝数的增多使长链脂肪酸含量明显升高[4]。……
