药学学报, 2013, 48(6): 953-959
引用本文:
徐艳红, 杨 欣, 张 争, 梁 良, 魏建和. 白木香3-羟基-3-甲基戊二酰辅酶A还原酶基因 AsHMGR2的克隆及表达分析[J]. 药学学报, 2013, 48(6): 953-959.
XU Yan-hong, YANG Xin, ZHANG Zheng, LIANG Liang, WEI Jian-he. Cloning and expression analysis of HMG-CoA reductase from Aquilaria sinensis (Lour.) Gilg[J]. Acta Pharmaceutica Sinica, 2013, 48(6): 953-959.

白木香3-羟基-3-甲基戊二酰辅酶A还原酶基因 AsHMGR2的克隆及表达分析
徐艳红1, 杨 欣1, 张 争1, 2, 梁 良1, 魏建和1, 2*
(1. 中国医学科学院、北京协和医学院药用植物研究所, 北京 100193; 2. 中国医学科学院、北京协和医学院药用植物研究所海南分所, 海南省南药资源保护与开发重点实验室, 海南 万宁 571533)
摘要:

3-羟基-3-甲基戊二酰辅酶A还原酶 (HMGR) 是萜类合成甲羟戊酸 (MVA)途径中的第一个关键限速酶。本研究根据课题组白木香转录组数据库中的HMGR2部分转录本序列设计引物, 利用RT-PCRRACE技术克隆HMGR2基因的全长cDNA序列, 并对其进行生物信息学分析; 利用荧光定量PCR检测AsHMGR2基因在不同组织和不同伤害处理下的表达差异。克隆得到的白木香AsHMGR2基因开放阅读框为1 749 bp, 编码582个氨基酸, GenBank登录号为KC140287。组织表达分析的结果显示, AsHMGR2基因主要在根和茎尖中表达, 其次是主干, 叶中的表达量最低; 该基因同时受物理伤害和结香液处理的诱导, 分别在6 h8 h达到最高表达水平。本研究通过AsHMGR2基因的全长cDNA克隆和表达特性分析, 为后续深入研究其在沉香倍半萜合成途径的功能奠定了基础。

关键词:   
Cloning and expression analysis of HMG-CoA reductase from Aquilaria sinensis (Lour.) Gilg
Abstract:

3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) is the first rate-limiting enzyme for sesquiterpene synthesis in the mevalonate (MVA) pathway.  The specific primers were designed according to the transcript sequence of AsHMGR2 from the Aquilaria sinensis (Lour.) Gilg transcriptome database.  The full- length cDNA of AsHMGR2 was cloned by RT-PCR and rapid amplification of cDNA ends (RACE) technology, and was analyzed at bioinformatics levels; AsHMGR2 expression profiles in different tissues and in responds to different treatments were analyzed by real-time PCR.  The length of AsHMGR2 Open Reading Frame (ORF) was 1 749 bp, encoding 582 amino acids.  The GenBank accession number is KC140287.  Tissue expression analysis indicated that AsHMGR2 was mainly expressed in root and shoot tips, followed by stem, and was lowest in leaves.  Inducible-experiments showed that the genes were induced by mechanical wound as well as chemical liquid induction, and reached the highest expression level at 6 h and 8 h, separately.  The full-length cDNA of AsHMGR2 and its expression patterns will provide a foundation for further research on its function in agarwood sesquiterpene biosynthesis.

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