YNB without ammonium sulphate, without potassium

Company: MP Biomedicals

Catalog#: 114029622

In vitro Assays to Evaluate Specificity and Affinity in Protein-phospholipid Interactions

Hendry Susila, Snježana Jurić, Katarzyna Gawarecka, Kyung Sook Chung, Suhyun Jin, Soo-Jin Kim, Zeeshan Nasim, Geummin Youn and Ji Hoon Ahn

Published online: 2022-05-20

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Mating Based Split-ubiquitin Assay for Detection of Protein Interactions

Author:

Wijitra Horaruang and Ben Zhang,

Date:

2017-05-05

[Abstract] The mating based split-ubiquitin (mbSUS) assay is an alternative method to the classical yeast two-hybrid system with a number of advantages. The mbSUS assay relies on the ubiquitin-degradation pathway as a sensor for protein-protein interactions, and it is suitable for the determination of interactions between full-length proteins that are cytosolic or membrane-bound. Here we describe the mbSUS assay protocol which has been used for detecting the interaction between K⁺ channel and SNARE proteins (Grefen et al., 2010 and 2015; Zhang et al., 2015 and 2016) [摘要] 基于交配的分ubiquitin（mbSUS）测定是具有许多优点的经典酵母双杂交系统的替代方法。 mbSUS测定依赖于泛素降解途径作为蛋白质 - 蛋白质相互作用的传感器，并且它适用于测定细胞溶质或膜结合的全长蛋白质之间的相互作用。在这里，我们描述了已经用于检测K ⁺通道和SNARE蛋白之间的相互作用的mbSUS测定方案（Grefen等人，2010和2015; Zhang 等等，2015和2016）

背景图1是mbSUS测定的概况。泛素部分被分成两半，N末端半突变（NubG）以避免重组。泛素部分（Cub）的C末端一半与转录报告基因复合物PLV（Protein A-LexA-VP16）连接。两种蛋白质（X和Y）分别与NubG和CubPLV融合产生蛋白质 - 蛋白质相互作用分析系统。转化后，酵母菌株THY.AP5含有NubG-X融合蛋白，而酵母菌株THY.AP4含有Y-CubPLV融合蛋白。在交配后，在二倍体酵母中，如果蛋白质X和Y彼此相互作用，则将重新组装功能性泛素，这导致PLV的切割。释放的转录蛋白复合物PLV可以开启报告基因（ADE2，HIS3），并允许酵母生长在选择性培养基上。

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YNB without ammonium sulphate, without potassium

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