Abstract:We study the target search and cleavage mechanisms of type II restriction endonucleases. In our single molecule technique, micron sized beads are tethered with single DNAs in a microfluidic flow cell and imaged using video microscopy. Under low protein concentration, the cleavage time is limited by the target search time. Using this technique, we have demonstrated that the target search strategy of NdeI employs a single 1D scan of 1000bp DNA at low salt concentration, but switches to a search mode requiring mu… Show more
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