蛋白质-蛋白质、蛋白质-核酸相互作用研究方法及在人肠道病毒A71型研究中的应用

Research Methods in Protein-Protein and Protein-Nucleic Acid Interactions and Application in the Study of Human Enterovirus A71

研究蛋白质-蛋白质相互作用的方法主要有酵母双杂交、噬菌体展示、免疫共沉淀、谷光苷肽巯基转移酶沉淀、细胞内共定位、亲和印迹、病毒铺覆蛋白结合技术、表面等离子共振、荧光共振能量转移等技术。检测蛋白质-核酸相互作用的方法主要包括酵母单杂交、染色质免疫沉淀、电泳迁移率实验、DNA-蛋白质印迹、报告基因、免疫共沉淀、谷光苷肽巯基转移酶沉淀、噬菌体展示等技术。在我们实验室对人肠道病毒A71型(EV-A71)的研究中经常会用到这些方法,但在该研究领域尚未有中文综述发表,因此本文对EV-A71研究中这些方法的应用进行了综述,该综述对蛋白质-蛋白质、蛋白质-核酸在其他病毒研究中的应用同样具有重要启示。

Methods for analyses of protein-protein interactions include： yeast two hybrid （Y2H）, phage dis- play （PD）, co-immunoprecipitation （Co-IP）, glutathione S-transferase pull-down （GST pull-down）, cellu- lar co-localization, far-western blotting, virus overlay protein binding assay （VOPBA）, surface plasmon resonance （SPR）, and fluorescence resonance energy transfer （FRET）. Technologies for the detection of protein-nucleic acid interactions include： yeast one hybrid （Y1H）, chromatin immunoprecipitation （CHIP）, electrophoretic mobility shift assay （EMSA）, Southwestern blotting, reporter gene, Co-IP, GST pull-down, and PD. These methods are often used in the study of the human enterovirus A71 （EV-A71） by our research team. Reviews in the Chinese literature in this field are lacking, so we reviewed applica- tions of these methods in the study of EV-A71. This review may impart important knowledge in the re- search of other viruses with regard to protein-protein and protein-nucleic acid interactions.