Development of a novel active targeting system based on split SNAP-tag

本实验开发了一种基于分裂SNAP标签的新型主动靶向系统。首先，研究了分裂SNAP标签互补。将SNAP标签片段融合到蛋白质中，使其能够相互作用。与荧光SNAP标签底物孵育后，表达分裂SNAP标签融合蛋白的细胞在这些蛋白相互作用时产生荧光信号。接下来，我们利用噬菌体PhiX 174的Gene-A* 蛋白（Gene-A*）对单链DNA（ssDNA）进行了位点特异性标记，并将其应用于蛋白质间相互作用的研究。

In this experiment, a novel active targeting system based on split SNAP-tag was developed. Firstly, split SNAP-tag complementation was studied. Split SNAP-tags, fragments of divided SNAP-tag were fused to proteins that can interact with each other. After incubation with a fluorescent SNAP-tag substrate, cells that expressed split SNAP-tag fusion proteins generated fluorescent signals when these proteins interacted. It was shown that split SNAP-tag labeling method should be a useful tool for visualization of protein-protein interaction processes.Next, for using DNA aptamer in our system, we developed a method for site-specific labeling of single-stranded DNA (ssDNA) to a recombinant protein of interest (POI) through the Gene-A* protein (Gene-A*) from bacteriophage phi X174, without any chemical modifications of ssDNA.