Xu Yan-Hong, Yang Xin, Zhang Zheng, Liang Liang, Wei Jian-He
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.
Yao Xue Xue Bao. 2013 Jun;48(6):953-9.
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.
3-羟基-3-甲基戊二酰辅酶A还原酶（HMGR）是甲羟戊酸（MVA）途径中倍半萜合成的首个限速酶。根据白木香转录组数据库中AsHMGR2的转录本序列设计特异性引物。通过RT-PCR和cDNA末端快速扩增（RACE）技术克隆AsHMGR2的全长cDNA，并进行生物信息学分析；通过实时PCR分析AsHMGR2在不同组织中的表达谱以及对不同处理的响应。AsHMGR2开放阅读框（ORF）长度为1749 bp，编码582个氨基酸。GenBank登录号为KC140287。组织表达分析表明，AsHMGR2主要在根尖和茎尖表达，其次是茎，在叶中表达量最低。诱导实验表明，该基因受机械损伤和化学液体诱导，分别在6 h和8 h达到最高表达水平。AsHMGR2全长cDNA及其表达模式将为进一步研究其在沉香倍半萜生物合成中的功能奠定基础。
