Citation: Yun Fei BAI, Qin Yu GE, Tong Xiang LI, Jin Ke WANG, Quan Jun LIU, Zu Hong LU. Endonuclease-based Method for Detecting the Sequence Specific DNA Binding Protein on Double-stranded DNA Microarray[J]. Chinese Chemical Letters, ;2005, 16(5): 651-654. shu

Endonuclease-based Method for Detecting the Sequence Specific DNA Binding Protein on Double-stranded DNA Microarray

Chien-Shiung Wu Laboratory, Department of Biological Science and Medical Engineering, Southeast University, Nanjing 210096

Received Date: 20 May 2004

Fund Project: This work was supported by the projects 30371606 and 60341002 of the National Natural Science Foundation of China, and also of the grant 2002AA2Z2041 from the National High Tech Program of China.

The double-stranded DNA (dsDNA) probe contains two different protein binding sites.One is for DNA-binding proteins to be detected and the other is for a DNA restriction enzyme.The two sites were arranged together with no base interval. The working principle of the capturing dsDNA probe is described as follows:the capturing probe can be cut with the DNA restriction enzyme (such as EcoR I) to cause a sticky terminal, if the probe is not bound with a target protein, and the sticky terminal can be extended and labeled with Cy3-dUTP by DNA polymerase. When the probe is bound with a target protein, the probe is not capable to be cut by the restriction enzyme because of space obstruction. The amount of the target DNA binding proteins can be measured according to the variations of fluorescent signals of the corresponding probes.
- Double stranded DNA microarray,
- DNA binding protein,
- label-free detection
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