Abstract
Panax notoginseng has been extensively used as a traditional Chinese medicine. In the current study, molecular cloning and characterization of PnbHLH1 transcription factor were explored in Panax notoginseng. The full length of the PnbHLH1 gene obtained by splicing was 1430 bp, encoding 321 amino acids. Prokaryotic expression vector pET-28a-PnbHLH1 was constructed and transferred into the BL21 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 PnbHLH1 transgenic cells were higher than those in the wild cells. Similarly, the total saponin contents were increased in the PnbHLH1 transgenic cell lines compared with the wild cell lines. Such results suggest that the PnbHLH1 transcription factor is a positive regulator in the biosynthesis of triterpenoid saponins in Panax notoginseng.
Highlights
IntroductionP. notoginseng saponins (PNS) are triterpenoids mainly synthesized through the mevalonic acid (MVA)
All the results indicated that PnbHLH1 transcription factors (TFs) could bind with the E-box cis-acting element
The positive were designed according to the hygromycin resistance gene (HPT) on T-DNA for screening the rate pCAMBIA1300s-PnbHLH1 vector was transferred into P. notoginseng cells and 16 anti-Hyg cell lines positive transgenic by PCR. cell
Summary
P. notoginseng saponins (PNS) are triterpenoids mainly synthesized through the mevalonic acid (MVA). Transcription factors are DNA binding proteins which bind to the promoter region of genes to regulate gene expression [3]. The operating transcription factor is an efficient strategy to control the biosynthetic pathway because of its unique multi-point regulation. With the elucidation of the secondary metabolic pathway of plants and regulatory mechanisms, especially the identification of transcription factors regulating the biosynthesis of specific secondary metabolites, gene engineering based on transcription factors has been proven to have strong effects on improving the yields of target metabolites [4]. The basic amino acid region is located at the N-terminus of the bHLH transcription factor which has a binding site and can identify DNA [6].
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
