红外荧光成像技术分析DNA-蛋白复合物电泳迁移率的变动  

An infrared imaging system for detecting electrophoretic mobility shift of DNA-protein complexes

作　　者：王毅飞[1] 蔡德鸿[1] 陈宏[1] 莫永炎[2] 伊娜[3] 邢飞跃[4] 

机构地区：[1]南方医科大学珠江医院内分泌科,广东广州510282 [2]南方医科大学病理生理学教研室,广东广州510515 [3]广州市中医院内分泌科,广东广州510130 [4]暨南大学组织移植与免疫中心,广东广州510632

出　　处：《南方医科大学学报》2009年第2期289-291,共3页Journal of Southern Medical University

摘　　要：目的通过红外荧光技术研究糖皮质激素诱导亮氨酸拉链蛋白(GILZ)与过氧化物酶体增殖物活化受体γ2(PPARγ2)启动子寡核苷酸的结合,了解一种非放射性研究电泳迁移率变动分析的新方法。方法原核表达制备GILZ蛋白,与红外荧光染料IRDye800标记的PPARγ2启动子寡核苷酸结合,丙烯酰胺非变性胶电泳分离,Odyssey红外荧光成像系统扫描分析。结果一条DNA-蛋白结合复合物图像清晰直观,信号强度随着蛋白量的增加而增加。结论红外荧光技术可用于DNA-蛋白复合物电泳迁移率变动的分析,具有灵敏度高、操作方便的优点。Objective To establish a new non-radioactive method for electrophoretic mobility shift assay （EMSA） to investigate the binding between glucocorticoid induced leucine zipper （GILZ） and peroxisome proliferator-activated receptor-gamma 2 （PPARγ2） promoter oligonucleotides. Methods GILZ protein prepared by prokaryotic expression was linked to PPARγ2 promoter oligonucleotides end-labeled with IRDye 800 infrared dye. The DNA-protein complex was separated with non-denatured polyacrylamide gel and scanned with the Odyssey. Infrared Imaging System. Results One lane of DNA-protein complex was clearly presented, and the signal intensity increased along with the increment of the protein load. Conclusion This infrared imaging system can be used for EMSA for detecting the DNA-protein complex with high sensitivity efficiency and allows easy operation.

关 键 词：红外荧光 电泳迁移率 DNA-蛋白复合物 糖皮质激素诱导亮氨酸拉链蛋白 过氧化物酶体增殖物活化受体γ2 

分 类 号：Q51-33[生物学—生物化学]