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利用 RecA 蛋白对 DNA 支架进行纳米级可编程的序列特异性图案化。

Nanoscale programmable sequence-specific patterning of DNA scaffolds using RecA protein.

机构信息

School of Electronic and Electrical Engineering, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, UK.

出版信息

Nanotechnology. 2012 Sep 14;23(36):365301. doi: 10.1088/0957-4484/23/36/365301. Epub 2012 Aug 21.

Abstract

Molecular self-assembly inherent to many biological molecules, in conjunction with suitable molecular scaffolds to facilitate programmable positioning of nanoscale objects, offers a promising approach for the integration of functional nanoscale complexes into macroscopic host devices. Here, we report the use of the protein RecA as a means of highly efficient programmable patterning of double-stranded (ds)DNA molecules with molecular-scale precision at specific locations along the DNA strand. RecA proteins form nucleoprotein filaments with single-stranded (ss)DNA molecules, which are chosen to be of sequence homologous to the desired binding region on the dsDNA scaffold. We show that the patterning yield can be in excess of 85% and we demonstrate that concurrent patterning of multiple locations on the same dsDNA scaffold can be achieved with separation between the assembled nucleoprotein filaments of less than 4 nm. This is an important prerequisite for this programmable and flexible DNA scaffold patterning technique to be employed in molecular- and nanoscale assembly applications.

摘要

许多生物分子固有的分子自组装,与合适的分子支架相结合,有利于纳米级物体的可编程定位,为将功能纳米级复合物集成到宏观主体设备中提供了一种很有前途的方法。在这里,我们报告了使用 RecA 蛋白作为一种手段,以分子级精度在双链 (ds)DNA 分子的特定位置上进行高效的可编程图案化,这些位置沿着 DNA 链选择。RecA 蛋白与单链 (ss)DNA 分子形成核蛋白丝,这些 ssDNA 分子的序列与 dsDNA 支架上所需的结合区域同源。我们表明,图案化的产率可以超过 85%,并且我们证明可以在相同的 dsDNA 支架上的多个位置同时进行图案化,组装的核蛋白丝之间的间隔小于 4nm。这是将这种可编程和灵活的 DNA 支架图案化技术应用于分子和纳米级组装应用的重要前提。

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