Citation: Min Li Li, Dong Rui Zhou, Hong Zhao, Jin Ke Wang, Zu Hong Lu. Endonuclease-rolling circle amplification-based method for sensitive analysis of DNA-binding protein[J]. Chinese Chemical Letters, ;2009, 20(11): 1315-1318. doi: 10.1016/j.cclet.2009.06.008 shu

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

a The State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China;
b Key Laboratory of Child Development and Learning Science, Ministry of Education, Southeast University, Nanjing 210096, China

Received Date: 31 March 2009

Fund Project: 60701008 and 60771024).This research is supported by the National Natural Science Foundation of China (Nos. 60501010

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.
- Rolling circle amplification,
- DNA-binding protein,
- Microarray
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