Flow dichroism of DNA: A new apparatus and further studies

✍ Scribed by Patrik R. Callis; Norman Davidson

• Book ID: 102762235
• Publisher: Wiley (John Wiley & Sons)
• Year: 1969
• Tongue: English
• Weight: 937 KB
• Volume: 7
• Category: Article
• ISSN: 0006-3525
• DOI: 10.1002/bip.1969.360070306

✦ Synopsis

A new apparatus for the study of flow dichroism of macromolecules is described. The flow is down a long, narrow channel and an unpolarised light beam propagates along the flow direction. For a molecule such as DNA, in which the transition moments of the chromophores are perpendicular to the axis of orientation, an increase of absorbance is observed during flow. The apparatus is best suited for macromolecules which are readily orientable or a t high shear gradients so that the extinction angle is close to 0". The apparatus has the following advantages: dilute macromolecule solutions can be used; high shear gradients are easily obtained; only small volumes of solution are needed. The flow can be stopped rapidly so that relaxation times for disorientation can be studied. The flow dichorism of native, two-stranded DNA has been measured for the molecular weight range of 0.6 X 106 to 125 X 108, and for the shear gradient range (in aqueous solution a t 25OC) from 200 sec-l to 21000 sec-l. At a fked gradient the dichroism increases with molecular weight, but the curve is concave downwards. At a given molecular weight the dichroism increases with increasing shear gradient, but the curve is concave downwards. When the solvent viscosity and temperature are varied, the dichroism is a function of q ( G ) / T showing that the orientation is due to hydrodynamic shear stress and that the flexibility of DNA in a flow field is not due to local denaturation. The Zimm-Rouse theory with no parameters taken from flow optical data predicts the correct order of magnitude of the dichroism but the experimentally observed shear gradient and molecular weight dependence do not fit the theory. This is an expected result, since the theory is believed to be applicable only a t small distortions and extensions of the macromolecule.