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

机译:使用RecA蛋白的DNA支架的纳米级可编程序列特异性模式

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摘要

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 4nm. This is an important prerequisite for this programmable and flexible DNA scaffold patterning technique to be employed in molecular-and nanoscale assembly applications.
机译:许多生物分子固有的分子自组装,再加上合适的分子支架以促进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宿主设备中提供了一种有前途的方法。在这里,我们报告蛋白质RecA的使用作为一种高效的双链(ds)DNA分子的可编程模式的手段,该分子在沿DNA链的特定位置具有分子尺度的精度。 RecA蛋白与单链(ss)DNA分子形成核蛋白丝,其选择的序列与dsDNA支架上所需的结合区同源。我们显示出图案产率可以超过85%,并且我们证明在组装的dsDNA支架上的小于4nm的核蛋白细丝之间的分离可以实现同一dsDNA支架上多个位置的同时图案化。这是在分子和纳米级组装应用中采用这种可编程和灵活的DNA支架构图技术的重要先决条件。

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