Vanderperre Solène, Duffraisse Marilyne, Place Christophe, Merabet Samir
Institut de Génomique Fonctionnelle de Lyon (IGFL), UMR5242, Ecole Normale Supérieure de Lyon (ENSL), CNRS, Université de Lyon, Lyon, France.
FLPENS, CNRS UMR572, Ecole Normale Supérieure de Lyon (ENSL), Université de Lyon, Lyon, France.
Methods Mol Biol. 2025;2889:39-51. doi: 10.1007/978-1-0716-4322-8_4.
Bimolecular Fluorescence Complementation (BiFC) is a powerful molecular imaging method used to visualize protein-protein interactions (PPIs) in living cells or organisms. BiFC is based on the reassociation of hemi-fragments of a monomeric fluorescent protein upon spatial proximity. It is compatible with conventional light microscopy, providing a resolution that is constrained by the diffraction of light to around 250 nm. PAmCherry1, a fluorescent protein compatible for BiFC and Photoactivated Localization Microscopy (PALM), allows the analysis of PPIs with nanometer spatial resolution and single molecule sensitivity. To date, this so-called BiFC-PALM approach has only been described in human cell culture. Here, we present a protocol for performing BiFC-PALM in Drosophila larval salivary glands, using the interaction between the Hox protein Ultrabithorax (Ubx) and the generic Hox cofactor Extradenticle (Exd) as a model system.
双分子荧光互补（BiFC）是一种强大的分子成像方法，用于在活细胞或生物体中可视化蛋白质-蛋白质相互作用（PPI）。BiFC基于单体荧光蛋白的半片段在空间接近时的重新结合。它与传统光学显微镜兼容，提供的分辨率受光的衍射限制，约为250纳米。PAmCherry1是一种与BiFC和光激活定位显微镜（PALM）兼容的荧光蛋白，可实现具有纳米空间分辨率和单分子灵敏度的PPI分析。迄今为止，这种所谓的BiFC-PALM方法仅在人类细胞培养中有所描述。在此，我们展示了一种在果蝇幼虫唾液腺中进行BiFC-PALM的方案，使用同源异型蛋白超双胸（Ubx）与通用同源异型辅因子额外齿（Exd）之间的相互作用作为模型系统。
