利用生物传感器实时监测T7 RNA聚合酶与偶氮苯连接的T7启动子之间的相互作用。

Real time monitoring of the interaction of T7 RNA polymerase with azobenzene-tethered T7 promoter by biosensor.

作者信息

Liu Mingzhe, Asanuma Hiroyuki, Komiyama Makoto

机构信息

Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.

出版信息

Nucleic Acids Symp Ser (Oxf). 2004(48):221-2. doi: 10.1093/nass/48.1.221.

DOI:10.1093/nass/48.1.221

PMID:17150558

Abstract

We have already reported that transcription reaction by phage T7 or SP6 RNA polymerase (RNAP) can be reversibly photo-regulated with azobenzene-tethered promoter. Transcription reaction proceeded faster by UV irradiation than visible light irradiation. In the present study, binding of T7 RNAP to its azobenzene-tethered promoter was directly monitored by use of affinity biosensor. When azobenzene-tethered T7 promoter was immobilized on the sensor surface, response based on the binding of RNAP increased by UV light irradiation rather than by visible light irradiation. Thus, photo-regulation of transcription reaction by azobenzene-tethered promoter was attributed to the change of binding property of RNAP to the promoter by trans-cis isomerization of azobenzene.

摘要

我们已经报道过，噬菌体T7或SP6 RNA聚合酶（RNAP）的转录反应可以通过偶氮苯连接的启动子进行可逆的光调控。紫外线照射下的转录反应比可见光照射下进行得更快。在本研究中，通过使用亲和生物传感器直接监测T7 RNAP与其偶氮苯连接的启动子的结合。当偶氮苯连接的T7启动子固定在传感器表面时，基于RNAP结合的响应在紫外线照射下增加，而不是在可见光照射下增加。因此，偶氮苯连接的启动子对转录反应的光调控归因于偶氮苯的反式-顺式异构化导致RNAP与启动子结合特性的改变。