| Endonuclease-based Method for Detecting the Sequence Specific DNA Binding Protein on Double-stranded DNA Microarray |
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| 作者姓名： | YunFeiBAI QinYuGE TongXiangLI JinKeWANG QuanJunLIU ZuHongLU |
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| 作者单位： | Chien-ShiungWuLaboratory,DepartmentofBiologicalScienceandMedicalEngineering.SoutheastUniversity,Nanjing210096 |
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| 摘 要： | 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.
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| 关 键 词： | 核酸内切酶 探测方法 DNA 序列 蛋白质 微阵列 化学结构 |
Endonuclease-based Method for Detecting the Sequence Specific DNA Binding Protein on Double-stranded DNA Microarray |
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| YunFeiBAI QinYuGE TongXiangLI JinKeWANG QuanJunLIU ZuHongLU.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. |
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| Authors: | Yun Fei BAI Qin Yu GE Tong Xiang LI Jin Ke WANG Quan Jun LIU Zu Hong LU Chien-Shiung Wu Laboratory |
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| Institution: | Yun Fei BAI,Qin Yu GE,Tong Xiang LI,Jin Ke WANG,Quan Jun LIU,Zu Hong LU Chien-Shiung Wu Laboratory,Department of Biological Science and Medical Engineering,Southeast University,Nanjing 210096 |
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| Abstract: | 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. |
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| Keywords: | Double stranded DNA microarray DNA binding protein label-free detection |
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