Stains Cliff I, Porter Jason R, Ooi Aik T, Segal David J, Ghosh Indraneel
Department of Chemistry and Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ 85721, USA.
J Am Chem Soc. 2005 Aug 10;127(31):10782-3. doi: 10.1021/ja051969w.
We describe a general methodology for the direct detection of DNA by the design of a split-protein system that reassembles to form an active complex only in the presence of a targeted DNA sequence. This approach, called SEquence Enabled Reassembly (SEER) of proteins, combines the ability to rationally dissect proteins to construct oligomerization-dependent protein reassembly systems and the availability of DNA binding Cys2-His2 zinc-finger motifs for the recognition of specific DNA sequences. We demonstrate the feasibility of the SEER approach utilizing the split green fluorescent protein appended to appropriate zinc fingers, such that chromophore formation is only catalyzed in the presence of DNA sequences that incorporate binding sites for both zinc fingers.
我们描述了一种通过设计一种分裂蛋白系统直接检测DNA的通用方法，该系统仅在存在靶向DNA序列时重新组装形成活性复合物。这种方法称为蛋白质的序列驱动重组装（SEER），它结合了合理剖析蛋白质以构建依赖寡聚化的蛋白质重组装系统的能力，以及用于识别特定DNA序列的DNA结合Cys2-His2锌指基序的可用性。我们利用连接到适当锌指上的分裂绿色荧光蛋白证明了SEER方法的可行性，使得发色团的形成仅在包含两个锌指结合位点的DNA序列存在时才被催化。
