Liang Liang, Han Xiao-Min, Zhang Zheng, Guo Qing-Mei, Xu Yan-Hong, Liu Juan, Liao Yong-Cui
Zhongguo Zhong Yao Za Zhi. 2014 May;39(10):1767-71.
The study aimed to clone the open reading frame of cinnamate 4-hydroxylase (C4H) from Aquilaria sinensis and analyze the bioinformatics and expression of the gene. One unique sequence containing C4H domain was discovered in our previous reported wound transcriptome dataset of A. sinensis. The open reading frame of C4H was cloned by RT-PCR strategy with the template of mixed RNA extracted from A. sinensis stem which treated by different wound time. The bioinformatic analysis of this gene and its corresponding protein was performed. C4H expression profiles in responds to MeJA (methyl jasmonate) application were analyzed by real-time PCR. The length of C4H open reading frame (ORF) was 1 515 bp, encoding 514 amino acids. The GenBank accession number is KF134783. Inducible-experiments showed that the genes were induced by mechanical wound as well as MeJA induction, and reached the highest expression level at 8 h and 20 h, respectively. The full-length cDNA of C4H and its expression patterns will provide a foundation for further research on its function in the molecular mechanisms of aromatic compounds and flavonoids biosynthesis.
该研究旨在克隆白木香肉桂酸-4-羟化酶（C4H）的开放阅读框，并分析该基因的生物信息学及表达情况。在我们之前报道的白木香创伤转录组数据集中发现了一个包含C4H结构域的独特序列。以不同创伤时间处理的白木香茎中提取的混合RNA为模板，通过RT-PCR策略克隆了C4H的开放阅读框。对该基因及其相应蛋白质进行了生物信息学分析。通过实时PCR分析了C4H在茉莉酸甲酯（MeJA）处理下的表达谱。C4H开放阅读框（ORF）长度为1515 bp，编码514个氨基酸。GenBank登录号为KF134783。诱导实验表明，该基因受机械创伤和MeJA诱导，分别在8 h和20 h达到最高表达水平。C4H全长cDNA及其表达模式将为进一步研究其在芳香族化合物和黄酮类生物合成分子机制中的功能奠定基础。
