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Activity and function studies of the promoter cis-acting elements of the key enzymes in saponins biosynthesis of DS from Panax notoginseng
Protoplasma ( IF 2.9 ) Pub Date : 2021-05-02 , DOI: 10.1007/s00709-021-01653-x
Yujie Zheng 1 , Rongyu Shao 1 , Pengguo Xia 1 , Zongsuo Liang 1 , Kaijing Yan 2
Affiliation  

Abstract

Panax notoginseng is a traditional Chinese medicine for the treatment of blood diseases, in which saponins were the main active components. Dammarenediol synthase (DS) is a key enzyme in the saponin synthesis pathway of P. notoginseng. The promoter is an important region to regulate gene expression, and the study of the promoter sequence provides important evidence for revealing the mechanism of gene expression regulation. However, there was still little research on the promoter function of P. notoginseng. In this study, the 1382 bp promoter upstream of DS from P. notoginseng was cloned and sequenced. The promoter sequence was analyzed by online databases. The plant expression vector fused with the β-glucuronidase gene was constructed and transferred into Agrobacterium tumefaciens. Then tobacco was injected, and its response to exogenous hormones (gibberellin and abscisic acid) was studied by transient expression to verify its unique action elements. The results showed that the tobacco leaves transferred with DS promoter had significantly increased GUS protease activity after spraying GA and ABA, indicating that both DS promoter can specifically and significantly respond to exogenous GA and ABA signal. These findings will help us to better understand the regulatory mechanisms of the upstream region of the DS gene and provide a basis for future research on the interaction of cis-acting elements of promoters with related transcription factors.

Graphical abstract



中文翻译:

三七DS皂苷生物合成关键酶启动子顺式作用元件的活性和功能研究

摘要

三七是一种治疗血液病的中药,其中皂甙为主要活性成分。Dammarenediol synthase (DS) 是三七皂苷合成途径中的关键酶。启动子是调控基因表达的重要区域,启动子序列的研究为揭示基因表达调控机制提供了重要证据。然而,关于三七的启动子功能的研究还很少。在本研究中,三七DS上游的 1382 bp 启动子被克隆和测序。通过在线数据库分析启动子序列。构建了融合了β-葡萄糖醛酸酶基因的植物表达载体,并将其转移到根癌农杆菌中。然后注入烟草,并通过瞬时表达研究其对外源激素(赤霉素和脱落酸)的反应,以验证其独特的作用元件。结果表明,转染DS启动子的烟叶喷施GA和ABA后,GUS蛋白酶活性显着提高,说明DS启动子均能特异性且显着响应外源GA和ABA信号。这些发现将有助于我们更好地了解DS上游区域的调控机制 为今后研究启动子顺式作用元件与相关转录因子的相互作用提供依据。

图形概要

更新日期:2021-05-02
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