Tu Li-Chan, Zhang Yi-Feng, Su Ping, Hu Tian-Yuan, Tong Yu-Ru, Guan Hong-Yu, Zhao Yu-Jun, Zhang Xia-Nan, Yuan Yuan, Gao Wei, Huang Lu-Qi
School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2017 Jan;42(2):220-225. doi: 10.19540/j.cnki.cjcmm.20161222.042.
Based on the transcriptome data, the study cloned full-length cDNA of TwGPPS1 and TwGPPS2 genes from Tripterygium wilfordii suspension cells and then analyzed the bioinformation of the sequence and protein expression. The cloned TwGPPS1 has a 1 278 bp open reading frame (ORF) encoding a polypeptide of 425 amino acids. The deduced isoelectric point (pI) was 6.68, a calculated molecular weight was about 47.189 kDa. The full-length cDNA of the TwGPPS2 contains a 1 269 bp open reading frame (ORF) encoding a polypeptide of 422 amino acids. The deduced isoelectric point (pI) was 6.71, a calculated molecular weight was about 46.774 kDa.The entire reading frame of TwGPPS1,2 was cloned into the pET-32a(+) vector and expressed in E. coli BL21 (DE3) cells to obtain the TwGPPS protein, which laid a basis for further study on the regulation of terpenoid secondary metabolism and biological synthesis.
基于转录组数据，该研究从雷公藤悬浮细胞中克隆了TwGPPS1和TwGPPS2基因的全长cDNA，然后分析了序列的生物信息和蛋白质表达。克隆得到的TwGPPS1具有一个1278 bp的开放阅读框（ORF），编码一个由425个氨基酸组成的多肽。推导的等电点（pI）为6.68，计算得到的分子量约为47.189 kDa。TwGPPS2的全长cDNA包含一个1269 bp的开放阅读框（ORF），编码一个由422个氨基酸组成的多肽。推导的等电点（pI）为6.71，计算得到的分子量约为46.774 kDa。将TwGPPS1、2的整个阅读框克隆到pET-32a(+)载体中，并在大肠杆菌BL21(DE3)细胞中表达以获得TwGPPS蛋白，这为进一步研究萜类次生代谢调控和生物合成奠定了基础。
