Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, Beijing 100084, China.
Int J Mol Sci. 2019 Apr 14;20(8):1843. doi: 10.3390/ijms20081843.
Paclitaxel is an important anticancer drug. The phytohormone jasmonic acid can significantly induce the biosynthesis of paclitaxel in , but the molecular mechanism has not yet been resolved. To establish the jasmonic acid signalling pathway of , based on the gene of the jasmonic acid signalling pathway of , sequence analysis was performed to isolate the jasmonic acid signal from the transcriptome, a transcriptional cluster of pathway gene homologs and the full length of 22 genes were obtained by RACE PCR at 5' and 3': two EI ubiquitin ligase genes, COI1-1 and COI1-2;7 MYC bHLH type transcription factor (MYC2, MYC3, MYC4, JAM1, JAM2, EGL3, TT8); 12 JAZ genes containing the ZIM domain; and MED25, one of the components of the transcriptional complex. The protein interaction between each were confirmed by yeast two hybridization and bimolecular fluorescence complementation based on similar genes interaction in Arabidopsis. A similar jasmonate signaling pathway was illustrated in . All known paclitaxel biosynthesis genes promoters were isolated by genome walker PCR. To investigate the jasmonate signaling effect on these genes' expression, the transcription activity of MYC2, MYC3 and MYC4 on these promoters were examined. There are 12, 10 and 11 paclitaxel biosynthesis genes promoters that could be activated by MYC2, MYC3 and MYC4.
紫杉醇是一种重要的抗癌药物。植物激素茉莉酸可以显著诱导合成紫杉醇，但分子机制尚未解决。为了建立的茉莉酸信号通路，根据茉莉酸信号通路的基因，对转录组进行了序列分析，以分离出茉莉酸信号，通过 RACE-PCR 获得了途径基因同源物的转录簇和全长 22 个基因的 5'和 3'：两个 EI 泛素连接酶基因 COI1-1 和 COI1-2；7 个 MYC bHLH 转录因子（MYC2、MYC3、MYC4、JAM1、JAM2、EGL3、TT8）；12 个含有 ZIM 结构域的 JAZ 基因；以及转录复合物的一个组成部分 MED25。通过酵母双杂交和双分子荧光互补实验，基于拟南芥中类似基因的相互作用，证实了它们之间的蛋白质相互作用。在中说明了一个类似的茉莉酸信号通路。通过基因组步移 PCR 分离了所有已知的紫杉醇生物合成基因启动子。为了研究茉莉酸信号对这些基因表达的影响，检测了 MYC2、MYC3 和 MYC4 对这些启动子的转录活性。有 12、10 和 11 个紫杉醇生物合成基因启动子可以被 MYC2、MYC3 和 MYC4 激活。
