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Science 8 November 1985:
Vol. 230. no. 4726, pp. 679 - 681
DOI: 10.1126/science.2996145
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Articles

Science, Vol 230, Issue 4726, 679-681
Copyright © 1985 by American Association for the Advancement of Science

articles

Iron(II) EDTA used to measure the helical twist along any DNA molecule

TD Tullius and BA Dombroski

A new method has been devised to measure the number of base pairs per helical turn along any DNA molecule in solution. A DNA restriction fragment is adsorbed onto crystalline calcium phosphate, fragmented by reaction with iron(II) EDTA, and subjected to electrophoresis on a denaturing polyacrylamide gel. A modulated cutting pattern results, which gives directly the helical periodicity of the DNA molecule. A 150-base pair sequence directly upstream of the thymidine kinase gene of the type 1 herpes simplex virus was found to have an overall helical twist of 10.5 base pairs per turn, which is characteristic of the B conformation of DNA. In addition, purines 3' to pyrimidines showed lower than expected reactivity toward the iron cutting reagent, which is evidence for sequence-dependent variability in DNA conformation.


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