沙棘苹果酸代谢关键基因ME的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of Key Gene ME in Malic Acid Metabolism of Sea Buckthorn

作　　者：黄瑶 李贺 包博闻 黄兰 黄佳妮 范宇畅 Huang Yao;Li He;Bao Bowen;Huang Lan;Huang Jiani;Fan Yuchang(College of Environment and Resources,Dalian Minzu University,Dalian,116600)

机构地区：[1]大连民族大学环境与资源学院,大连116600

出　　处：《分子植物育种》2022年第5期1470-1476,共7页Molecular Plant Breeding

基　　金：大连民族大学大学生创新创业训练计划项目(201912026174);辽宁省自然科学基金计划项目(2020-MZLH-09)共同资助。

摘　　要：苹果酸酶(ME)参与苹果酸的降解,是植物苹果酸代谢的关键酶。本研究根据前期的沙棘转录组测序数据设计特异性引物,利用RT-PCR技术克隆ME基因(NAD-ME,NADP-ME)并进行生物信息学分析。结果表明,沙棘NAD-ME基因的完整开放阅读框长1902 bp,编码一个由634个氨基酸组成的亲水性蛋白,主要由α-螺旋(44.16%)、无规则卷曲(39.91%)和少量β-折叠(15.93%)构成,与枣的NAD-ME同源性最高。NADP-ME的完整开放阅读框长1926 bp,编码一个由642个氨基酸组成的亲水性蛋白,主要由α-螺旋(37.07%)、无规则卷曲(45.64%)和少量β-折叠(15.29%)构成,与川桑和核桃NADP-ME的同源性较高。研究结果为探索沙棘苹果酸代谢的分子调控机制奠定理论基础。Malice enzymes(ME)are key enzymes in malic acid metabolism in plant,which involved in the degradation of malic acid.In this study,the specific primers were designed based on the sea buckthorn transcriptome sequencing data.Afterwards ME genes(NAD-ME,NADP-ME)were cloned using RT-PCR method and bioinformatics analysis were carried out.The ORF(open reading frame)of NAD-ME was 1902 bp length,encoding a hydrophilic protein of 634 amino acids.It was mainly composed ofα-helix(44.16%),random coil(39.91%)and a small amount ofβ-sheet(15.93%),and had close phylogenetic relationships with Zizyphus jujuba.The ORF of NADP-ME was 1926 bp length,encoding a hydrophilic protein of 642 amino acids.It mainly composed ofα-helix(37.07%),random coil(45.64%)and a small amount ofβ-sheet(15.29%),and had close phylogenetic relationships with mulberry.These results provide a foundation to explore the molecular regulatory mechanism of malate metabolism in sea buckthorn.

关 键 词：沙棘 苹果酸酶 克隆 生物信息学分析 

分 类 号：S793.6[农业科学—林木遗传育种]