摘要
4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,CMK)是植物萜类化合物生物合成途径甲基赤藓醇-4-磷酸途径(methylerythritol-4-phosphate pathway,DXP/MEP)上的第4个关键酶。该研究根据樟树转录组数据,利用RT-PCR(reverse transcription-PCR)方法从药用植物樟树Cinnamomum camphora中克隆得到4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶基因,命名为Cc CMK1(Gene Bank登录号Ku376098),对其序列进行生物信息学分析,结果表明该基因开放阅读框(ORF)为1 212 bp,编码403个氨基酸,推测其相对分子质量为44.46 k Da,等电点(theoretical p I)为4.99。跨膜结构分析表明CMK蛋白不存在跨膜结构。信号肽分析表明其为非分泌蛋白,不存在信号肽。亚细胞定位表明其定位于叶绿体中。利用荧光实时PCR检测了龙脑樟、油樟、芳樟、脑樟和异樟5种化学型樟树中Cc CMK1基因的表达量,结果表明Cc CMK1基因在油樟中的表达量最高,龙脑樟中表达量最低,为樟树萜类化合物生物合成途径关键酶基因元件挖掘提供研究基础。
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 Cc CMK1,then deposited it in Gene Bank（ Accession number： Ku376098）. Bioinformatics analysis showed the open reading frame（ ORF） of the Cc CMK1 was 1 212 bp. The putative protein encoded 403 amino acids,and its molecular weight was 44. 46 k Da 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 Cc CMK1 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.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2016年第9期1578-1584,共7页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(30901963)
