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Nucleic Acids Research, 1991, Vol. 19, No. 24 6811-6817
© 1991

MOLECULAR BIOLOGY

Probing of DNA structure with osmium tetroxide,2,2'-bipridine. Adduct-specific antibodies

Alena Kuderovà-Krecovà, Alexander M. Poverenny1 and Emil Palecek*

Institute of Biophysics, Czechoslovak Academy of Sciences Kralovopolska 135, 612 65 Brno, Czechoslovakia 1nstitute of Medical Radiology 149 029 Obninsk, USSR

*To whom correspondence should be addressed

Received September 26, 1991. Accepted November 19, 1991.

Antibodies against DNA modified with a single-strand selective probe, OsO4 in complex with 2,2'-bipyridine (Os,bipy), were raised in rabbits. These antibodies were fractionated using affinity column chromatography and fractions S89-II and S89-III characterized as highly specific for DNA-Os,bipy adduct with no cross reactivity to at least 1000-fold excess of unmodified DNA, RNA and Os.blpy-modified and unmodified proteins. Crossreactivity to Os,bipy-modified RNA was very small. S89-II showed no cross-reactivity to DNA modified with OsO4 completed with fetramethylethylenediamine or with bathophenanthroline disulphonic acid and to DNA oxidized with KWJnO4. It cross-reacted, however, with DNA modified with OsO4,1,10-phenanthroline complex. The limit of detection of Immunodot-blot analysis of extensively Os.bipy-modified DNA was below 0.5 pg. Small extent of Os,bipy-modificaf ion of supercoiled and linearized plasmids can be detected by DNA gel retardation and immunoblotting techniques. E. coli cells contain DNA regions in which bases are accessible to the single-strand selective probe.


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Home page
 J. Cell Sci.Home page
E Palecek, M Robert-Nicoud, and T. Jovin
Local opening of the DNA double helix in eukaryotic cells detected by osmium probe and adduct-specific immunofluorescence
J. Cell Sci., January 3, 1993; 104(3): 653 - 661.
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