Reichman Pavel, Dissmeyer Nico
Independent Junior Research Group on Protein Recognition and Degradation, Leibniz Institute of Plant Biochemistry (IPB) and Science Campus Halle - Plant-Based Bioeconomy, Halle (Saale), Germany.
Methods Mol Biol. 2017;1669:387-406. doi: 10.1007/978-1-4939-7286-9_29.
Determination of the general capacity of proteolytic activity of a certain cell or tissue type can be crucial for an assessment of various features of an organism's growth and development and also for the optimization of biotechnological applications. Here, we describe the use of chimeric protein stability reporters that can be detected by standard laboratory techniques such as histological staining, selection using selective media or fluorescence microscopy. Dependent on the expression of the reporters due to the promoters applied, cell- and tissue-specific questions can be addressed. Here, we concentrate on methods which can be used for large-scale screening for protein stability changes rather than for detailed protein stability studies.
确定特定细胞或组织类型的蛋白水解活性的总体能力,对于评估生物体生长和发育的各种特征以及优化生物技术应用可能至关重要。在此,我们描述了嵌合蛋白稳定性报告基因的用途,这些报告基因可通过标准实验室技术检测,如组织学染色、使用选择性培养基进行筛选或荧光显微镜观察。根据所应用启动子导致的报告基因表达情况,可以解决细胞和组织特异性问题。在此,我们专注于可用于大规模筛选蛋白质稳定性变化而非详细蛋白质稳定性研究的方法。