Endonuclease-rolling circle amplification-based method for sensitive analysis of DNA-binding protein
Endonuclease-rolling circle amplification-based method for sensitive analysis of DNA-binding protein
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基于核酸内切酶滚环扩增的 DNA 结合蛋白灵敏分析方法
DOI：
10.1016/j.cclet.2009.06.008
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发表时间：
2009-11
影响因子：
9.1
通讯作者：
Zhao, Hong
中科院分区：
文献类型：
--
作者：
Wang, Jin Ke;Zhou, Dong Rui;Lu, Zu Hong;Li, Min Li;Zhao, Hong
建立了一种灵敏的定性检测微阵列上DNA结合蛋白的方法。将其中由生物素引物和环状单链DNA（ssDNA）形成部分双链体区域的DNA复合物点样在微阵列上。核酸内切酶识别位点（ERS）和DNA结合位点（DBS）并排排列在双链体区域内。该检测系统的工作原理是：当DNA结合蛋白捕获DBS时，核酸内切酶不能与ERS结合，DNA复合物中的固定化引物通过滚环扩增（RCA）沿着环状ssDNA延伸。当没有蛋白质保护DBS时，ERS可被内切酶攻击，随后不发生滚环扩增。因此，由于RCA的信号放大，我们可以高灵敏度地检测序列特异性DNA结合活性。
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.
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影响因子：
14.9
作者：
Wang J;Li T;Guo X;Lu Z
通讯作者：
Lu Z
影响因子：
48
作者：
Jarvius, Jonas;Melin, Jonas;Nilsson, Mats
通讯作者：
Nilsson, Mats
影响因子：
46.9
作者：
Schweitzer, B;Roberts, S;Kingsmore, SF
通讯作者：
Kingsmore, SF
影响因子：
30.8
作者：
Mukherjee, S;Berger, MF;Bulyk, ML
通讯作者：
Bulyk, ML
影响因子：
46.9
作者：
Heyduk, T;Heyduk, E
通讯作者：
Heyduk, E