葫芦科PAL基因家族生物信息学分析及甜瓜PAL4基因克隆
Bioinformatics Analysis of PAL Gene Family in Cucurbitaceae and the PAL4 Gene Cloning in Cucumis melo L.
孙宇蛟 1陈欣 1崔浩楠 2吴川 2刘识 2栾非时 3王学征2
作者信息
- 1. 东北地区园艺作物生物学与种质创制农业部重点实验室, 哈尔滨, 150030;东北农业大学生命科学学院,哈尔滨, 150030
- 2. 东北地区园艺作物生物学与种质创制农业部重点实验室, 哈尔滨, 150030;东北农业大学园艺园林学院, 哈尔滨, 150030
- 3. 东北地区园艺作物生物学与种质创制农业部重点实验室, 哈尔滨, 150030;东北农业大学生命科学学院,哈尔滨, 150030;东北农业大学园艺园林学院, 哈尔滨, 150030
- 折叠
摘要
苯丙氨酸解氨酶是苯丙烷代谢途径的第一个酶,对植物发育、抗逆及信号传导起着尤为重要的作用.为了在葫芦科作物中更为深入地研究苯丙氨酸解氨酶的结构及理化性质、发掘更多潜在的苯丙氨酸解氨酶基因,本研究从拟南芥(Arabidopsis thaliana)4个苯丙氨酸解氨酶基因(At-PAL)入手,通过BLAST比对方法在葫芦科数据库(http://cucurbitgenomics.org/)中鉴定甜瓜(Cucumis melo L.)、西瓜(Citrullus lanatus)、黄瓜(Cucumis sativus L.)苯丙氨酸解氨酶的同源基因,并进行其生物信息学预测分析.结果表明:在甜瓜、西瓜、黄瓜中分别比对出4条、6条、5条同源基因,分别命名为MELO-PAL 1～4、Cla-PAL 1～6、Csa-PAL 1～5.其各级结构及理化性质与拟南芥苯丙氨酸解氨酶基因均较为相似.并对它们进行了系统进化树的构建,结果表明各物种间苯丙氨酸解氨酶基因家族间同源性较高,并利用分子生物学的手段,克隆了甜瓜的第四条比对基因(MELO-PAL4),将其连入T载体中进行测序.测序结果显示,克隆的基因序列与比对序列间总体相似度为92.54％.本研究为进一步发掘葫芦科作物PAL基因功能提供了重要的帮助.
Abstract
Phenylalanine ammonia-lyase is the first entry enzyme in the metabolism of phenylpropanoid, which plays a key role in plant development, stress resistance and signal transduction. In order to further study the structure and physicochemical properties of phenylalanine ammonia-lyase in Cucurbitaceae, and explore more potential phenylalanine ammonia lyase genes, this study started from four L-phenylalanine ammonia-lyase genes in Arabidopsis thaliana, and the homologous genes of melon (Cucumis melo L.), watermelon (Citrullus lanatus) and cucumber (Cucumis sativus L.) were identified by BLAST comparison in Cucurbitaceae database (http://cucurbitgenomics.org/), and their bioinformatics prediction and analysis were carried out. The results showed that 4, 6 and 5 homologous genes were identified in melon, watermelon and cucumber, respectively, which were named as MELO-PAL 1～4, Cla-PAL 1～6, and Csa-PAL 1～5. Its structure and physicochemical properties at different levels were all relatively similar to those of Arabidopsis thaliana. Phylogenetic tree analysis indicated that phenylalanine ammonia-lyasegenes from each species shared highly homology. In addition, MELO-PAL4 was cloned using molecular biology, which was subsequently ligated to T vector for sequencing. The sequencing results implied that the total similarity between cloned gene sequences and BLAST one was 92.54％. This study provided important help for the further excavation of the functions of PAL gene in Cucurbitaceae.
关键词
苯丙氨酸解氨酶/葫芦科/二代测序/生物信息学分析/基因克隆Key words
L-phenylalanine ammonia-lyase/Cucurbitaceae/Second generation sequencing/Bioinformatics analysis/Gene cloning引用本文复制引用
基金项目
国家自然科学基金(31372088)
国家西甜瓜产业技术体系岗位专家项目(CARS-026-02)
出版年
2018
NETL
NSTL
维普
万方数据