Jing Li, Zheng Han, Yao Na, Chen Mei-Lan, Shen Ye, Huang Lu-Qi
State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
AnHui University of Chinese Medicine, Hefei 230038, China.
Zhongguo Zhong Yao Za Zhi. 2016 May;41(9):1578-1584. doi: 10.4268/cjcmm20160902.
The 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase was the fourth key enzymes in plant terpenoid biosynthesis pathway of methyl erythritol phosphate pathway(MEP). According to the study of Cinnamomum camphora transcriptome data,we abtained the 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase gene using RT-PCR,and named CcCMK1,then deposited it in GeneBank(Accession number： Ku376098).Bioinformatics analysis showed the open reading frame (ORF) of the CcCMK1 was 1 212 bp.The putative protein encoded 403 amino acids,and its molecular weight was 44.46 kDa and theoretically isoelectric point was 4.99.Transmembrane structure analysis showed that there was no transmembrane structure. Signal peptide analysis showed that it was a non secretory protein, and there was no signal peptide. The subcellular localization showed that the chloroplast was located in the chloroplast.Analysis of the expression of CcCMK1 gene in five chemotypes of C. camphora using Real-time PCR showed its expression level was highest in C. longepaniculatum, and the lowest in Borneol camphor.This research provided a basis for characterizing the key enzyme genes of terpenoid biosynthetic pathway in C. camphora.
4-二磷酸胞苷-2-C-甲基-D-赤藓糖醇激酶是植物甲基赤藓糖醇磷酸途径（MEP）萜类生物合成途径中的第四个关键酶。根据对樟树转录组数据的研究，我们利用RT-PCR获得了4-二磷酸胞苷-2-C-甲基-D-赤藓糖醇激酶基因，并将其命名为CcCMK1，然后将其存入基因库（登录号：Ku376098）。生物信息学分析表明，CcCMK1的开放阅读框（ORF）为1212 bp。推测的蛋白质编码403个氨基酸，分子量为44.46 kDa，理论等电点为4.99。跨膜结构分析表明不存在跨膜结构。信号肽分析表明它是一种非分泌蛋白，不存在信号肽。亚细胞定位表明其位于叶绿体中。利用实时荧光定量PCR分析CcCMK1基因在樟树五种化学型中的表达情况，结果表明其在长柄樟中的表达水平最高，在龙脑樟中的表达水平最低。本研究为阐明樟树萜类生物合成途径关键酶基因提供了依据。
