The interaction of DNA restriction fragments with strong electric fields will be investigated using the technique of transient electric birefringence. Reversing field experiments will demonstrate whether the DNA molecules undergo a progressive change in conformation with increasing field strength. Studies of the Kerr constant as a function of counterion concentration and type will provide a test of the Manning theory of counterion condensation, as well as the Mandel and O'Konski theories of electric polarizability. Studies of the Kerr constant of very small fragments will show whether the recently observed L2 dependence of the electric polarizability (and hence the induced dipole) on length extends to fragments smaller than 100 base pairs. Birefringence saturation curves will be measured as a function of counterion concentration and type. The results will be correlated with the birefringence decay curves in order to interpret the apparent linear dependence of the birefringence on electric field strength in intermediate field strength regions. Intercalation of dye molecules into the DNA helix will be investigated with monodisperse fragments, in order to show whether or not the DNA helix is actually elongated by intercalation. Finally, the effect of electric fields on the thermal denaturation of DNA will be studied. The results of all these experiments will clarify the behavior of DNA molecules in strong electric fields, and thus can identify the electrical forces acting on the DNA molecule in the cell; the electrical potential across a typical cell membrane is of the same order of magnitude as the fields used in electric birefringence experiments. The long range goals of this research are to understand the physical properties of DNA and to determine the electrical forces acting on polyelectrolytes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM029690-06
Application #
3277316
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1981-07-01
Project End
1987-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
6
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Iowa
Department
Type
Schools of Medicine
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242
Stellwagen, Nancy C; Stellwagen, Earle (2009) Effect of the matrix on DNA electrophoretic mobility. J Chromatogr A 1216:1917-29
Stellwagen, Nancy C (2009) Electrophoresis of DNA in agarose gels, polyacrylamide gels and in free solution. Electrophoresis 30 Suppl 1:S188-95
Stellwagen, Nancy C (2006) Curved DNA molecules migrate anomalously slowly in polyacrylamide gels even at zero gel concentration. Electrophoresis 27:1163-8
Lu, Yongjun; Weers, Brock D; Stellwagen, Nancy C (2005) Intrinsic curvature in the VP1 gene of SV40: comparison of solution and gel results. Biophys J 88:1191-206
Lu, Yongjun; Weers, Brock D; Stellwagen, Nancy C (2003) Analysis of DNA bending by transient electric birefringence. Biopolymers 70:270-88
Lu, Yongjun; Weers, Brock D; Stellwagen, Nancy C (2003) Analysis of the intrinsic bend in the M13 origin of replication by atomic force microscopy. Biophys J 85:409-15
Stellwagen, Earle; Lu, Yongjun; Stellwagen, Nancy C (2003) Unified description of electrophoresis and diffusion for DNA and other polyions. Biochemistry 42:11745-50
Stellwagen, Nancy C (2003) Sequence-dependent bending in plasmid pUC19. Electrophoresis 24:3467-75
Stellwagen, Earle; Stellwagen, Nancy C (2003) Probing the electrostatic shielding of DNA with capillary electrophoresis. Biophys J 84:1855-66
Stellwagen, Earle; Stellwagen, Nancy C (2002) The free solution mobility of DNA in Tris-acetate-EDTA buffers of different concentrations, with and without added NaCl. Electrophoresis 23:1935-41

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