Chapter 06 磁镊介绍和应用.pdfVIP

Chapter 06 Introduction and applications Smith et al., Science, 1992 Strick et al., Science, 1996 Magnetic tweezers cartoon Magnetic tweezers cartoon The tightest non-covalent bond known roughly ~100kT Biotin + SA ↔ Biotin-SA Kassoc = [B-SA]/ [B][SA] = 1014M-1 The complex is also extremely stable over a wide range of temperature and pH. 254 AA = 46 x 93 x 104 Å = 4 x 15kD =60kD /stenkamp/stefanieweb/abstract.html /tutorial/streptavidin/index.html Magnetic Traps DNA twisting σ Antibody-ligand Antibody Magnetic Traps: Measuring DNA stretch Diffraction rings Focal point Z Z Magnetic Traps: Measuring DNA stretch Magnetic Tweezers and DNA Can be conveniently used to stretch and twist DNA. With Super-paramagnetic bead, no permanent dipole. It is the gradient of the force, which determines the direction, the force is up. (i.e. where B is highest) • DNA tends to be stretched out if move magnet up. • DNA also tends to twist if twist magnets (either mechanically, or electrically move magnets) Forces ranging from a few fN to nearly 100 pN: Huge Range Watch as a function of protein which interacts with DNA (polymerases, topoisomerases), as a function of chromatin: lo