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用于植物蛋白互作研究的酵母基于杂交的分裂泛素化系统

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The yeast mating-based Split-Ubiquitin System (mbSUS) used in the determination of protein-protein interaction in plant

摘要: 蛋白-蛋白相互作用对于植物细胞生理活动至关重要。为检测和确定两种蛋白之间的相互作用,许多不同的体内和体外方法已被发表。而酵母基于杂交的分裂泛素化(mbSUS)系统是经典的酵母双杂交技术替代方法,它不仅具有酵母双杂交快速便捷的优点,还弥补了传统酵母双杂交法在待测蛋白要求,假阳性率等方面的不足。目前,经过改进的mbSUS法可用于膜蛋白以及可溶性蛋白互作研究,并在植物中用于检测离子通道与膜转运蛋白的互作。本文对mbSUS法技术原理、实验方法以及实验注意事项进行详细描述,为植物蛋白互作研究奠定基础。
 
Abstract: The protein-protein interactions (PPIs) have a pivotal role in plant cells. There are numerous methods, in vivo or in vitro, have been developed to investigate the PPIs. The yeast mating-based Split-Ubiquitin System (mbSUS) is an alternative method of classic yeast-two-hybrid (Y2H) method. The mbSUS assay is not only as fast and inexpensive as the Y2H method, but also suitable for membrane proteins investigation with lower false positive rate. The recently improved mbSUS assay is suitable for the determination of PPIs among membrane proteins and soluble proteins, and successful used for detecting the interaction between ion channels and vesicle traffic proteins. Here, we describe the principles, protocols and experiment notes for mbSUS assay, providing an alternative method for PPI research in plants.
 

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[V1] 2023-10-23 11:03:34 ChinaXiv:202310.03415V1 下载全文
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