三七中PnbHLH1转录因子的分子克隆与特性分析
Molecular Cloning and Characterization of PnbHLH1 Transcription Factor in Panax notoginseng.
作者信息
Zhang Xiang, Ge Feng, Deng Bing, Shah Taif, Huang Zhuangjia, Liu Diqiu, Chen Chaoyin
机构信息
Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China.
出版信息
Molecules. 2017 Jul 29;22(8):1268. doi: 10.3390/molecules22081268.
has been extensively used as a traditional Chinese medicine. In the current study, molecular cloning and characterization of PnbHLH1 transcription factor were explored in . The full length of the gene obtained by splicing was 1430 bp, encoding 321 amino acids. Prokaryotic expression vector was constructed and transferred into the prokaryotic expression strain. An electrophoretic mobility shift assay of PnbHLH1 protein binding to E-box cis-acting elements verified that PnbHLH1 belonged to the bHLH class transcription factor which could interact with the promoter region of the E-box core sequence. The expression levels of key genes involved in the biosynthesis of triterpenoid saponins in transgenic cells were higher than those in the wild cells. Similarly, the total saponin contents were increased in the transgenic cell lines compared with the wild cell lines. Such results suggest that the transcription factor is a positive regulator in the biosynthesis of triterpenoid saponins in .
已被广泛用作传统中药。在当前研究中，对PnbHLH1转录因子进行了分子克隆和特性分析。通过拼接获得的该基因全长为1430 bp，编码321个氨基酸。构建了原核表达载体并将其转入原核表达菌株。PnbHLH1蛋白与E-box顺式作用元件结合的电泳迁移率变动分析证实，PnbHLH1属于bHLH类转录因子，可与E-box核心序列的启动子区域相互作用。转基因细胞中三萜皂苷生物合成相关关键基因的表达水平高于野生细胞。同样，与野生细胞系相比，转基因细胞系中的总皂苷含量增加。这些结果表明，该转录因子是三萜皂苷生物合成中的正调控因子。
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