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Subangstrom single-molecule measurements of motor proteins using a nanopore.

Nat. Biotechnol.. 2015;

DerringtonIan M，CraigJonathan M，StavaEric，LaszloAndrew H，RossBrian C，BrinkerhoffHenry，NovaIan C，DoeringKenji，TickmanBenjamin I，RonaghiMostafa，MandellJeffrey G，GundersonKevin L，GundlachJe

Products/Services Used	Details	Operation
Gene Synthesis	… Methods Proteins The M2-NNN-MspA protein 6 was custom ordered from GenScript Wild-type phi29 DNAP (833,000 U/ml; specific activity 83,000 U/mg) was obtained from Enzymatics or Epicenter Hel308 was expressed using standard techniques by in-house facilities …	Get A Quote

摘要

Techniques for measuring the motion of single motor proteins， such as FRET and optical tweezers， are limited to a resolution of ∼300 pm. We use ion current modulation through the protein nanopore MspA to observe translocation of helicase Hel308 on DNA with up to ∼40 pm sensitivity. This approach should be applicable to any protein that translocates on DNA or RNA， including helicases， polymerases， recombinases and DNA repair enzymes.