Endonuclease-rolling circle amplification-based method for sensitive analysis of DNA-binding protein

作     者：Min Li Li Dong Rui Zhou Hong Zhao Jin Ke Wang Zu Hong Lu 

作者机构：The State Key Laboratory of Bioelectronics Southeast University Nanjing 210096 China Key Laboratory of Child Development and Learning Science Ministry of Education Southeast University Nanjing 210096 China 

出 版 物：《Chinese Chemical Letters》 (中国化学快报（英文版）)

年 卷 期：2009年第20卷第11期

页      面：1315-1318页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 071009[理学-细胞生物学] 09[农学] 0703[理学-化学] 0901[农学-作物学] 090102[农学-作物遗传育种] 

基　　金：supported by the National Natural Science Foundation of China(Nos.60501010 60701008 and 60771024) 

主　　题：Rolling circle amplification DNA-binding protein Microarray 

摘      要：A sensitive approach for the qualitative detection of DNA-binding protein on the microarray was developed. DNA complexes in which a partial duplex region is formed from a biotin-primer and a circle single strand DNA （ssDNA） were spotted on a microarray. The endonuclease recognition site （ERS） and the DNA-binding sites （DBS） were arranged side by side within the duplex region. The working principle of the detection system is described as follows： when the DNA-binding protein capture the DBS, the endonuclease could not attach to the ERS, and the immobilized primer in the DNA complex could be extended along the circle ssDNA by rolling circle amplification （RCA）. When no protein protects the DBS, the ERS could be attacked by the endonuclease and subsequently no rolling circle amplification occurs. Thereby we can detect the sequence specific DNA-binding activity with high-sensitivity due to the signal amplification of RCA.