首页 | 本学科首页   官方微博 | 高级检索  
     检索      

苹果NADP依赖的苹果酸酶基因克隆、序列和表达分析
引用本文:董庆龙,王海荣,安淼,余贤美,王长君.苹果NADP依赖的苹果酸酶基因克隆、序列和表达分析[J].中国农业科学,2013,46(9):1857-1866.
作者姓名:董庆龙  王海荣  安淼  余贤美  王长君
作者单位:山东省果树研究所,山东泰安 271000
基金项目:山东省果树研究所所长科研基金(2012KY06)、国家自然科学基金项目(31100086)
摘    要:【目的】克隆苹果(Malus×domestica B.)中苹果酸代谢的关键酶基因MdNADP-ME,进行序列特征分析,研究MdNADP-ME在苹果中组织表达的情况。【方法】利用RT-PCR技术获得苹果MdNADP-ME1,2,3全长的cDNA序列。对该序列进行生物信息学分析,采用荧光实时定量PCR技术研究MdNADP-ME1,2,3的组织表达。【结果】测序结果显示,获得的3个NADP-苹果酸酶基因(MdNADP-ME1、MdNADP-ME2 和 MdNADP-ME3; GenBank 登录号为JX971883、JX971884和JX971885),其ORF分别为1 512、1 782和1 926 bp,推测其分别编码503、593和641个氨基酸的多肽。氨基酸序列和结构分析显示,这3个基因均含有5个保守的氨基酸区域(motif I-V),含有2个功能结构域:malic和NAD_bind_1_malic_enz。进化树分析结果显示,MdNADP-ME1 和MdNADP-ME2属于双子叶植物细胞质NADP-ME型,MdNADP-ME3属于双子叶植物质体NADP-ME型。荧光实时定量PCR结果表明,MdNADP-ME1-3在被检测的组织中均有表达,但表达差异明显。【结论】MdNADP-ME1-3属于植物NADP-ME家族,结构高度保守,并且在苹果的不同组织中有不同表达模式。

关 键 词:苹果    苹果酸酶    克隆    序列分析    表达分析
收稿时间:2012-10-23

Cloning, Sequence and Expression Analysis of NADP-Malic Enzyme Genes in Apple
DONG Qing-Long,WANG Hai-Rong,AN Miao,YU Xian-Mei,WANG Chang-Jun.Cloning, Sequence and Expression Analysis of NADP-Malic Enzyme Genes in Apple[J].Scientia Agricultura Sinica,2013,46(9):1857-1866.
Authors:DONG Qing-Long  WANG Hai-Rong  AN Miao  YU Xian-Mei  WANG Chang-Jun
Institution:Shandong Institute of Pomology, Tai’an 271000,Shandong
Abstract:【Objective】This study is aimed to characterize the genes of MdNADP-MEs involved in malic acid metabolism in Malus×domestica B. by gene cloning, sequence and expression of MdNADP-MEs in different tissues.【Method】 Three full-length cDNA sequences of MdNADP-ME1,2,3 were isolated by RT-PCR. The obtained cDNA sequences and the deduced amino acid sequences were analyzed with bioinformatics methods. qRT-PCR was used to assess the expression of MdNADP-MEs in different tissues.【Result】The sequencing results showed that three cDNAs (designated as MdNADP-ME1, 2 and 3; GenBank Accession No. JX971883, JX971884 and JX971885) were 1 512 bp, 1 782 bp and 1 926 bp. They contained an open reading frame (ORF) of 1 512 bp, 1 782 bp and 1 926 bp, respectively, and their ORFs encoded an protein with 503, 593 and 641 amino acids, respectively. Amino acid sequence and structure analysis indicated that MdNADP-MEs contained five conservative amino acid areas (motif I-V) and two functional structure domains: malic and NAD_bind_1_malic_enz. The result of phylogenetic analysis showed that MdNADP-ME1 and 2 belonged to cytosolic dicot NADP-ME (group I), MdNADP-ME3 belonged to plastidic dicot NADP-ME. qRT-PCR result showed that MdNADP-MEs were constitutively expressed in all examined tissues, but showed different expression levels.【Conclusion】MdNADP-ME1,2,3 belong to NADP-ME family, possess highly conserved structures, and show different expression patterns in different tissues of apple.
Keywords:apple  malic enzyme  cloning  sequence analysis  expression analysis
点击此处可从《中国农业科学》浏览原始摘要信息
点击此处可从《中国农业科学》下载免费的PDF全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号