Kumari Khushbu, Sherpa Tsheten, Dey Nrisingha
Division of Plant Biotechnology, Institute of Life Sciences, Bhubaneswar, Odisha, India.
Regional Centre for Biotechnology, National Capital Region Biotech Science Cluster, Faridabad, Haryana, India.
Front Plant Sci. 2024 Aug 6;15:1426479. doi: 10.3389/fpls.2024.1426479. eCollection 2024.
Promoters are one of the most important components for many gene-based research as they can fine-tune precise gene expression. Many unique plant promoters have been characterized, but strong promoters with dual expression in both monocot and dicot systems are still lacking. In this study, we attempted to make such a promoter by combining specific domains from monocot-infecting pararetroviral-based promoters sugarcane bacilliform virus (SCBV) and banana streak virus (BSV) to a strong dicot-infecting pararetroviral-based promoter mirabilis mosaic virus (MMV). The generated chimeric promoters, MS, SM, MB, and BM, were tested in monocot and dicot systems and further validated in transgenic tobacco plants. We found that the developed chimeric promoters were species-specific (monocot or dicot), which depended on their respective core promoter (CP) region. Furthermore, with this knowledge, deletion-hybrid promoters were developed and evaluated, which led to the development of a unique dual-expressing promoter, MSD3, with high gene expression efficiency (GUS and GFP reporter genes) in rice, pearl millet, and tobacco plants. We conclude that the MSD3 promoter can be an important genetic tool and will be valuable in plant biology research and application.
启动子是许多基于基因的研究中最重要的组成部分之一,因为它们可以精确调控基因表达。许多独特的植物启动子已被鉴定,但在单子叶和双子叶系统中都具有双重表达能力的强启动子仍然缺乏。在本研究中,我们尝试通过将来自感染单子叶的类逆转录病毒启动子甘蔗杆状病毒(SCBV)和香蕉条纹病毒(BSV)的特定结构域与感染双子叶的类逆转录病毒强启动子烟草花叶病毒(MMV)相结合来构建这样一个启动子。所产生的嵌合启动子MS、SM、MB和BM在单子叶和双子叶系统中进行了测试,并在转基因烟草植株中进一步验证。我们发现,所开发的嵌合启动子具有物种特异性(单子叶或双子叶),这取决于它们各自的核心启动子(CP)区域。此外,基于这一认识,我们开发并评估了缺失杂交启动子,从而开发出了一种独特的双重表达启动子MSD3,其在水稻、珍珠粟和烟草植株中具有高基因表达效率(GUS和GFP报告基因)。我们得出结论,MSD3启动子可以成为一种重要的遗传工具,在植物生物学研究和应用中具有重要价值。