Department of Rheumatology and Immunology, State Key Laboratory of Virology, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China.
Department of Pulmonary and Critical Care Medicine, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China.
Appl Microbiol Biotechnol. 2023 Jun;107(12):3955-3966. doi: 10.1007/s00253-023-12546-9. Epub 2023 May 11.
Fusion tag technology is an important tool for rapid separation, purification, and characterization of proteins. Combined with monoclonal antibodies, tag epitope systems can be rapidly adapted to many assay systems. A monoclonal antibody that reacts with the matrix protein of the rabies virus CVS-11 strain was reported. The epitope (termed M) targeted by this antibody contains only six amino acids. We examine whether this specific sequence epitope can be applied as a protein tag. We show ectopic expression of M-tagged proteins has little impact on cell viability or major signaling pathways. The M tag system can be used for western blotting, immunoprecipitation, immunofluorescence staining, and flow cytometry assays. The results indicate the specificity, sensitivity, and versatility of this novel epitope tag system are comparable to the widely used FLAG tag system, providing researchers with an additional tool for molecular analysis. KEY POINTS: • A short peptide (Pro Pro Tyr Asp Asp Asp) can be applied as a new tag. • The new epitope-tagging fusion system has no effect on the main cellular signaling pathway. • The epitope-tagging fusion system can be widely used for western blotting, immunoprecipitation, immunofluorescence, flow cytometry, etc.
融合标签技术是一种快速分离、纯化和鉴定蛋白质的重要工具。与单克隆抗体结合使用，标签表位系统可以快速适应许多检测系统。本文报道了一种与狂犬病病毒 CVS-11 株基质蛋白反应的单克隆抗体。该抗体的表位（称为 M）仅包含六个氨基酸。我们研究了这种特异性序列表位是否可作为蛋白质标签。结果表明，M 标签蛋白的异位表达对细胞活力或主要信号通路几乎没有影响。M 标签系统可用于 Western blot、免疫沉淀、免疫荧光染色和流式细胞术检测。结果表明，该新型表位标签系统的特异性、灵敏度和多功能性与广泛使用的 FLAG 标签系统相当，为研究人员提供了分子分析的另一种工具。关键点： • 短肽（Pro Pro Tyr Asp Asp Asp）可作为新标签应用。 • 新的表位标记融合系统对主要细胞信号通路没有影响。 • 表位标记融合系统可广泛用于 Western blot、免疫沉淀、免疫荧光、流式细胞术等。
