Tong Yu-ru, Su Ping, Zhang Meng, Zhao Yu-jun, Wang Xiu-juan, Gao Wei, Huang Lu-qi
Zhongguo Zhong Yao Za Zhi. 2015 Nov;40(22):4378-83.
To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
为了从雷公藤中克隆2-C-甲基-D-赤藓糖醇-4-磷酸胞苷转移酶（TwMCT）全长cDNA，根据转录组数据设计了特异性引物并进行了长链PCR。对TwMCT进行一系列生物信息学分析后，通过实时荧光定量聚合酶链反应（RT-qPCR）研究茉莉酸甲酯（MeJA）诱导的表达量。结果显示，TwMCT cDNA全长为1318个核苷酸，编码311个氨基酸。推导的TwMCT蛋白分子量约为34.14 kDa，理论等电点为8.65。RT-qPCR分析结果表明，用MeJA处理后，雷公藤悬浮细胞中TwMCT mRNA表达量升高，并在24小时达到最大值。从雷公藤中克隆和分析TwMCT基因，为研究次生代谢产物的功能和表达调控提供了基因元件。
