A Temperature-Jump Optical Trap for Single-Molecule Manipulation

被引：55

作者：

de Lorenzo, Sara ^{[1
,2
]}

Ribezzi-Crivellari, Marco ^{[1
]}

Ricardo Arias-Gonzalez, J. ^{[3
,4
]}

Smith, Steven B. ^{[5
]}

Ritort, Felix ^{[1
,2
]}

机构：

[1] Univ Barcelona, Dept Fis Fonamental, Barcelona, Spain

[2] Inst Salud Carlos III, Ciber BBN Bioingn Biomat & Nanomed, Madrid, Spain

[3] Inst Madrileno Estudios Avanzados Nanociencia IMD, Madrid, Spain

[4] CNB CSIC IMDEA Nanociencia Associated Unit Unidad, Madrid, Spain

[5] Steven B Smith Engn, Los Lunas, NM USA

来源：

BIOPHYSICAL JOURNAL | 2015年 / 108卷 / 12期

基金：

欧洲研究理事会;

关键词：

FORCE SPECTROSCOPY; STRANDED-DNA; TWEEZERS; DEPENDENCE; THERMODYNAMICS; MICROANALYSIS; STABILITY;

D O I：

10.1016/j.bpj.2015.05.017

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

To our knowledge, we have developed a novel temperature-jump optical tweezers setup that changes the temperature locally and rapidly. It uses a heating laser with a wavelength that is highly absorbed by water so it can cover a broad range of temperatures. This instrument can record several force-distance curves for one individual molecule at various temperatures with good thermal and mechanical stability. Our design has features to reduce convection and baseline shifts, which have troubled previous heating-laser instruments. As proof of accuracy, we used the instrument to carry out DNA unzipping experiments in which we derived the average basepair free energy, entropy, and enthalpy of formation of the DNA duplex in a range of temperatures between 5 degrees C and 50 degrees C. We also used the instrument to characterize the temperature-dependent elasticity of single-stranded DNA (ssDNA), where we find a significant condensation plateau at low force and low temperature. Oddly, the persistence length of ssDNA measured at high force seems to increase with temperature, contrary to simple entropic models.

引用

页码：2854 / 2864

页数：11

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