一个引物对所有:一个标准化的载体工具箱,用于蛋白质相互作用和植物蛋白质定位。

IF 6.3 1区 生物学 Q1 PLANT SCIENCES
Cailin Luo, Lihaitian Wang, Yuqin Wu, Jun Cai, Bin Peng, Jia Chen, Feng Yu, Dousheng Wu
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引用次数: 0

摘要

了解蛋白质-蛋白质相互作用(PPIs)对于阐明植物生长、发育和胁迫反应的分子机制至关重要。虽然分析PPI的技术已经发展得很好,但验证PPI通常需要四到五种技术的组合。对于每种技术,设计特定的引物将目标基因(GOI)克隆到给定的载体中。因此,验证PPI需要合成四到五个引物对,这使得该过程既复杂又昂贵。为了克服这些限制,我们优化并功能验证了一个基于in - fusion的矢量工具箱,用于在植物中灵活构建PPI矢量。标准化工具箱使得仅使用一对引物就可以将GOI克隆到不同的PPI载体中。此外,我们扩展了用于蛋白质表达和定位研究的标准化载体之一。使用相同的引物对,可以很容易地将GOI克隆到具有不同标签或荧光团的载体中。我们优化的载体工具箱将大大简化PPI载体以及其他植物表达载体的组装,用于亚细胞定位和蛋白质表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
One Primer Pair for All: A Standardized Vector Toolbox for Protein-Protein Interactions and Protein Localization in Plants.

Understanding protein-protein interactions (PPIs) is crucial for elucidating the molecular mechanisms underlying plant growth, development, and stress responses. While techniques for analyzing PPIs have been well developed, validating a PPI typically requires a combination of four to five techniques. For each technique, specific primers are designed to clone the gene of interest (GOI) into a given vector. As a result, validating a PPI requires the synthesis of four to five primer pairs, making the process both complex and costly. To overcome these limitations, we have optimized and functionally validated an In-Fusion-based vector toolbox for the flexible construction of PPI vectors in plants. The standardized toolbox facilitates the cloning of a GOI into different PPI vectors using only one primer pair. Furthermore, we extended one of the standardized vectors for protein expression and localization studies. With the same primer pair, a GOI can be easily cloned into vectors with different tags or fluorophores. Our optimized vector toolbox will significantly simplify the assembly of PPI vectors as well as other plant expression vectors for subcellular localization and protein expression.

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来源期刊
Plant, Cell & Environment
Plant, Cell & Environment 生物-植物科学
CiteScore
13.30
自引率
4.10%
发文量
253
审稿时长
1.8 months
期刊介绍: Plant, Cell & Environment is a premier plant science journal, offering valuable insights into plant responses to their environment. Committed to publishing high-quality theoretical and experimental research, the journal covers a broad spectrum of factors, spanning from molecular to community levels. Researchers exploring various aspects of plant biology, physiology, and ecology contribute to the journal's comprehensive understanding of plant-environment interactions.
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