Bhadouriya Sneha Lata, Karamchandani Arti Narendra, Nayak Namitha, Mehrotra Sandhya, Mehrotra Rajesh
Department of Biological Sciences, Birla Institute of Technology and Sciences Pilani, Goa campus, Goa, India.
Sci Rep. 2024 Jun 13;14(1):13657. doi: 10.1038/s41598-024-64537-z.
This work aimed to design a synthetic salt-inducible promoter using a cis-engineering approach. The designed promoter (PS) comprises a minimal promoter sequence for basal-level expression and upstream cis-regulatory elements (CREs) from promoters of salinity-stress-induced genes. The copy number, spacer lengths, and locations of CREs were manually determined based on their occurrence within native promoters. The initial activity profile of the synthesized PS promoter in transiently transformed N. tabacum leaves shows a seven-fold, five-fold, and four-fold increase in reporter GUS activity under salt, drought, and abscisic acid stress, respectively, at the 24-h interval, compared to the constitutive CaMV35S promoter. Analysis of gus expression in stable Arabidopsis transformants showed that the PS promoter induces over a two-fold increase in expression under drought or abscisic acid stress and a five-fold increase under salt stress at 24- and 48-h intervals, compared to the CaMV35S promoter. The promoter PS exhibits higher and more sustained activity under salt, drought, and abscisic acid stress compared to the constitutive CaMV35S.
这项工作旨在采用顺式工程方法设计一种合成的盐诱导型启动子。设计的启动子(PS)包含用于基础水平表达的最小启动子序列以及来自盐胁迫诱导基因启动子的上游顺式调控元件(CRE)。基于CRE在天然启动子中的出现情况,手动确定了其拷贝数、间隔长度和位置。在瞬时转化的烟草叶片中,合成的PS启动子的初始活性谱显示,与组成型CaMV35S启动子相比,在24小时间隔时,报告基因GUS活性在盐胁迫、干旱胁迫和脱落酸胁迫下分别增加了7倍、5倍和4倍。对稳定的拟南芥转化体中gus表达的分析表明,与CaMV35S启动子相比,PS启动子在24小时和48小时间隔时,在干旱或脱落酸胁迫下诱导表达增加两倍以上,在盐胁迫下诱导表达增加五倍。与组成型CaMV35S相比,启动子PS在盐胁迫、干旱胁迫和脱落酸胁迫下表现出更高且更持久的活性。