In vitro selection of DNA binding sites for transcription factor, PhaR, from Paracoccus denitrificans using genetic library on microbeads and flow cytometry
In vitro selection of DNA binding sites for transcription factor, PhaR, from Paracoccus denitrificans using genetic library on microbeads and flow cytometry
复制标题
DOI：
10.1263/jbb.101.440
复制
发表时间：
2006-05-01
影响因子：
2.8
通讯作者：
Nakano, Hideo
中科院分区：
文献类型：
--
作者：
Kojima, Takaaki;Yamane, Tsuneo;Nakano, Hideo
我们试图选择一个网站的DNA结合的转录因子，PhaR，从副球菌无细胞蛋白质合成表达，从微珠上的随机寡核苷酸文库，通过乳液PCR技术构建。PhaR是脱氮杆菌中的一种抑制蛋白，与phaP启动子区结合。利用该系统获得了三种PhaR结合DNA片段。所选片段含有一个完整的PhaR结合共有位点（TGC 1）、一个与TGC 1相似的序列和另一个PhaR结合位点（TGC 11）。
We attempted the selection of a site for DNA binding to a transcription factor, PhaR, from Paracoccus denitrificans expressed by cell-free protein synthesis, from a random oligonucleotide library on microbeads that was constructed by the emulsion PCR technique. PhaR is a repressor protein in P denitrificans that binds to the phaP promoter region. We acquired three types of PhaR-binding DNA fragment using this system. The selected fragments contained a perfect PhaR binding consensus site (TGC 1), a sequence similar to that of TGC 1, and another PhaR binding site (TGC 11).