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

在本实验中，开发了一种新型的基于分离式SNAP-Tag的主动目标系统。首先，研究了拆分式SNAP-Tag互补。拆分SNAP标签，将拆分的SNAP标签片段融合到能够相互作用的蛋白质中。在与荧光SNAP-Tag底物孵育后，表达分裂的SNAP-Tag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，为了在我们的系统中使用DNA适配子，我们发展了一种通过噬菌体Phi x174的基因-A*蛋白(gene-A*)将单链DNA(SsDNA)定位到重组蛋白(POI)的方法，而不需要对ssDNA进行任何化学修饰。

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.