GST-pulldown验证转录因子ZNF24与c-Myc的相互作用

摘要

Objective: In the previous yeast two-hybrid experiment, we used the SACN domain of transcription factor ZNF24 as bait and screened a clone including proto-oncogene c-Myc. To further verify the interaction of transcription factors ZNF24 and c-Myc. Methods: Through gene cloning and carrier construction technology ZNF24 and c-Myc were built into the different expression vector, the GST-pulldown technology was used to verify interactions of both expressed protein in vitro. Results: Electrophoresis appraisal and sequencing analysis showed that gene cloning right and carrier to build success, GST-pulldown successfully detected ZNF24 interact with c-Myc protein bands. Conclusion: GST-pulldown verified the interaction of ZNF24 and c-Myc in vitro.%目的：前期酵母双杂交实验中，我们以转录因子ZNF24的SCAN结构域为诱饵，筛选到的一个阳性克隆为原癌基因c-Myc，在此进一步验证ZNF24与c-Myc间的相互作用。方法：将ZNF24与c-Myc分别构建到pGEX-4T-2和pcDNA3.1表达载体上，利用GST-pulldown技术体外验证两者表达蛋白的相互作用。结果：电泳鉴定与测序分析表明目的基因克隆正确，载体构建成功；用GST-pulldown技术检测到ZNF24与c-Myc相互作用的蛋白条带。结论：GST-pulldown实验进一步表明ZNF24与c-Myc的相互作用，为验证ZNF24与c-Myc之间存在相互作用奠定了基础。