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Nucleic Acids Research, 2000, Vol. 28, No. 11 E51-e51
© 2000 Oxford University Press

Early melting of supercoiled DNA topoisomers observed by TGGE

Viktor Víglasky*, Marián Antalík1, Jozef Adamcík and Dusan Podhradsky

P.J. Safarik University, Faculty of Sciences, Department of Biochemistry, Moyzesova 11, 041 54 Koice, Slovakia and 1Institute of Experimental Physics, Slovak Academy of Sciences, Watsonova 47, 043 53 Koice, Slovakia

We have used temperature gradient gel electrophoresis (TGGE) to measure the progress of local denaturation in closed circular topoisomer DNA as a function of temperature and superhelicity ({sigma}). We describe the versatility of this method as a tool for detecting various conformational modifications of plasmid DNAs. The early melting temperature of a structural transition for any topoisomer is dependent on the value of superhelicity. Supercoiled topo­isomers represent a system of molecules that is sensitive to changes in temperature. We show that the topoisomer with the highest absolute value of superhelicity melts earlier than topoisomers with lower values. Thermal sensitivity of highly supercoiled plasmids could play a biologically important role in regulation of replication and expression in cells under thermal stress. The estimated melting temperature for plasmids with {sigma} < –0.05 is very significant because these temperatures for early melting are below physiological temperatures.

* To whom correspondence should be addressed. Tel: +421 95 62 235 82; Fax: +421 95 62 221 24; Email: viglasky@kosice.upjs.sk


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