Department of Physics and Astronomy, University of California, Los Angeles, Los Angeles, California 90095-1547, USA.
J Am Chem Soc. 2013 Aug 14;135(32):11879-86. doi: 10.1021/ja4043565. Epub 2013 Aug 1.
We report experiments where the activity of the enzyme luciferase from Renilla reniformis is controlled through a DNA spring attached to the enzyme. In the wake of previous work on kinases, these results establish that mechanical stress applied through the DNA springs is indeed a general method for the artificial control of enzymes, and for the quantitative study of mechano-chemical coupling in these molecules. We also show proof of concept of the luciferase construct as a sensitive molecular probe, detecting a specific DNA target sequence in an easy, one-step, homogeneous assay, as well as SNP detection without melting curve analysis.
我们报告了实验结果，其中通过连接到酶的 DNA 弹簧来控制来自肾形海肾的酶荧光素酶的活性。在先前关于激酶的工作之后，这些结果证实，通过 DNA 弹簧施加的机械应力确实是人工控制酶的通用方法，也是研究这些分子中机械化学耦联的定量方法。我们还展示了荧光素酶构建体作为灵敏分子探针的概念验证，该探针可以在简单的一步均相测定中检测到特定的 DNA 靶序列，并且无需进行熔融曲线分析即可检测 SNP。
