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水稻中编码3-羟基-3-甲基戊二酰辅酶A还原酶的Hmg2基因的分子特征分析

Molecular characterization of Hmg2 gene encoding a 3-hydroxy-methylglutaryl-CoA reductase in rice.

作者信息

Ha S H, Lee S W, Kim Y M, Hwang Y S

机构信息

Division of Biochemistry, National Institute of Agricultural Science and Technology, Rural Development Administration, Suwon, Korea.

出版信息

Mol Cells. 2001 Jun 30;11(3):295-302.

Abstract

Three genes encoding 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR, EC1.1.1.34), which converts HMG-CoA into mevalonate in the early key step of the plant isoprenoid pathway, were isolated by RT-PCR and rice cDNA and genomic library screening. A genomic Southern blot analysis confirmed that HMGR genes are present in three copies in rice. Of the three, the HMGR 2 gene (Hmg2) obtained as a cDNA clone and its genomic clone had 4 exons and 3 introns, and encoded a 576 amino acid peptide containing an open reading frame of 1,728 bp with a calculated Mw. of 61,150. The structure of rice Hmg2 had common features, based on its nucleotide and deduced amino acid sequence homologies, with other plant HMGR genes published to date. Rice Hmg2 transcripts were constitutively detected in all parts of the rice plant, except in lamina and their levels were high particularly in the leaf part of the dark-grown seedlings and mature flowers. Our result showed that mRNA levels of rice Hmg2 were strongly induced in seedlings and influorescence in the early development stage. Rice Hmg2 possibly has a housekeeping role involved in the sterol biosynthesis, among the possible roles of plant HMGR genes that have been suggested in other plants [Weissenborn et al. (1995)].

摘要

通过逆转录聚合酶链式反应(RT-PCR)以及水稻互补DNA(cDNA)和基因组文库筛选,分离出了三个编码3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR,EC1.1.1.34)的基因,该酶在植物类异戊二烯途径的早期关键步骤中将HMG-CoA转化为甲羟戊酸。基因组Southern杂交分析证实,HMGR基因在水稻中以三个拷贝存在。其中,作为cDNA克隆获得的HMGR 2基因(Hmg2)及其基因组克隆有4个外显子和3个内含子,编码一个含1728 bp开放阅读框的576个氨基酸的肽段,计算分子量为61,150。基于其核苷酸和推导的氨基酸序列同源性,水稻Hmg2的结构与迄今已发表的其他植物HMGR基因具有共同特征。除叶片外,在水稻植株的所有部位均能组成性检测到水稻Hmg2转录本,其水平在黑暗生长的幼苗和成熟花的叶片部分尤其高。我们的结果表明,水稻Hmg2的mRNA水平在早期发育阶段的幼苗和花序中受到强烈诱导。在其他植物中已提出的植物HMGR基因的可能作用中,水稻Hmg2可能在甾醇生物合成中起管家作用[魏森伯恩等人(1995年)]。

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