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  4. Measuring The Physical Step-Size Of Bacteriophage T7 Dna Helicase With Dual Optical Tweezers

Measuring The Physical Step-Size Of Bacteriophage T7 Dna Helicase With Dual Optical Tweezers

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bys2thesisPDF.pdf (15.07 MB)
Permanent Link(s)
https://hdl.handle.net/1813/29245
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Cornell Theses and Dissertations
Author
Smith, Benjamin
Abstract

Helicases are ubiquitous molecular motors which unwind double stranded DNA into two single strands. T7gp4 is a model hexameric helicase from the replication system of bacteriophage T7. The step size of T7gp4 was probed using single molecule optical trapping experiments. To obtain sufficient resolution over experimental timescales a dual optical trap was constructed, decoupling the experiment from the surface. Numerical simulations of helicase in the experimental configuration were conducted to examine the required measurement bandwidth. A novel method of forming double tethers between the dual traps was implemented. A range of steps of the helicase were observed. In combination with structural data and biochemical measurements, this agrees with a sequential mechanism for T7gp4 translocation.

Date Issued
2011-01-31
Keywords
single molecule biophysics
•
optical tweezers
•
helicase
Committee Chair
Wang, Michelle D
Committee Member
Sethna, James Patarasp
Xu, Chunhui
Degree Discipline
Physics
Degree Name
Ph. D., Physics
Degree Level
Doctor of Philosophy
Type
dissertation or thesis

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