Confirmation of replication interactome of TopBP1 using proximity-dependent biotinylation MS

Goal of the Experiment is to confirm the replication interactome of TopBP1 using stabile 293 FlpIn cell lines expressing TopBP1 adapter fused to APEX2. Proteins in its close proximity will become biotinylated by APEX2, thereby enabling the pulldown of biotinylated proteins and thus, replication involved substrates of TopBP1

本实验旨在验证TopBP1的复制相互作用组（replication interactome），采用稳定表达与抗坏血酸过氧化物酶2（APEX2）融合的TopBP1接头蛋白的293 FlpIn细胞系。定位于该融合蛋白邻近区域的蛋白质将被APEX2催化发生生物素化，借此可实现对生物素化蛋白的下拉实验（pulldown）富集，进而获取TopBP1的复制相关底物。