Plant Biotechnology Lab, Department of Botany, Faculty of Science, Dayalbagh Educational Institute (Deemed University), Dayalbagh, Agra, 282005, India.
Institute of Life Science, Nalco Square, Bhubaneshwar, Odisha, 751023, India.
Planta. 2021 Jan 21;253(2):42. doi: 10.1007/s00425-021-03565-9.
In this study, useful hybrid promoters were developed for efficient ectopic gene expression in monocot and dicot plants, and they hold strong prominence in both transgenic research and biotech industries. This study deals with developing novel synthetic promoters derived from Rice Tungro Bacilliform Virus (RTBV) and Mirabilis Mosaic Virus (MMV). Despite numerous availability, there is a severe scarcity of promoters universally suitable for monocot and dicot plants. Here, eight chimeric promoter constructs were synthesized as gBlocks gene fragments through domain swapping and hybridization by incorporating important domains of previously characterized RTBV and MMV promoters. The developed promoter constructs were assessed for transient GUS expression in tobacco protoplast (Xanthi Brad) and agro-infiltrated tobacco, petunia, rice and pearl millet. Protoplast expression analysis showed that two promoter constructs, namely pUPMA-RP1-MP1GUS and pUPMA-RP4-MP1GUS exhibited 3.56 and 2.5 times higher activities than that of the CaMV35S promoter. We had observed the similar type of expression patterns of these promoters in agroinfiltration-based transient studies. RP1-MP1 and RP4-MP1 promoters exhibited 1.87- and 1.68-fold increase expression in transgenic tobacco plants; while, a 1.95-fold increase was found in RP1-MP1 transgenic rice plants when compared their activities with CaMV35S promoter. Furthermore, on evaluating these promoter constructs for their expression in the bacterial system, pUPMA-RP1-MP1GFP was found to have the highest GFP expression. Moreover, the promoter construct was also evaluated for its capacity to express the HMP3 gene. Biobeads of encapsulated bacterial cells expressing HMP3 gene under control of the pUPMA-RP4-MP1 promoter were found to reduce 72.9% copper and 29.2% zinc concentration from wastewater. Our results had demonstrated that the developed promoter constructs could be used for translational research in dicot, monocot plants and bacterial systems for efficient gene expression.
在这项研究中，开发了有用的杂种启动子，用于在单子叶植物和双子叶植物中外源基因的高效表达，它们在转基因研究和生物技术产业中都具有重要地位。本研究涉及开发新型合成启动子，这些启动子源自水稻草丛斑驳病毒（RTBV）和美丽马兜铃花叶病毒（MMV）。尽管有许多启动子可用，但普遍适用于单子叶植物和双子叶植物的启动子仍然严重缺乏。在这里，通过域交换和杂交，合成了 8 个嵌合启动子构建体作为 gBlocks 基因片段，通过整合先前表征的 RTBV 和 MMV 启动子的重要结构域。通过瞬时转染烟草原生质体（Xanthi Brad）和农杆菌浸润烟草、矮牵牛、水稻和珍珠粟，评估了所开发的启动子构建体的 GUS 表达情况。原生质体表达分析表明，两个启动子构建体 pUPMA-RP1-MP1GUS 和 pUPMA-RP4-MP1GUS 的活性分别比 CaMV35S 启动子高 3.56 倍和 2.5 倍。我们在基于农杆菌浸润的瞬时研究中观察到了这些启动子类似的表达模式。RP1-MP1 和 RP4-MP1 启动子在转基因烟草植物中的表达分别增加了 1.87 倍和 1.68 倍；而在 RP1-MP1 转基因水稻植物中，与 CaMV35S 启动子相比，其活性增加了 1.95 倍。此外，在评估这些启动子构建体在细菌系统中的表达时，发现 pUPMA-RP1-MP1GFP 具有最高的 GFP 表达。此外，还评估了该启动子构建体表达 HMP3 基因的能力。在控制下表达 HMP3 基因的包埋细菌细胞的生物珠 pUPMA-RP4-MP1 发现可以将废水中的铜浓度降低 72.9%，锌浓度降低 29.2%。我们的研究结果表明，所开发的启动子构建体可用于翻译研究，在双子叶植物、单子叶植物和细菌系统中实现高效基因表达。
