Methods of detection of conformational change in a nucleic...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 21/00 (2006.01) G01N 21/00 (2006.01) G01N 33/50 (2006.01) G01N 33/52 (2006.01) G01N 33/53 (2006.01) G01N 33/566 (2006.01) G01N 33/84 (2006.01)

Patent

CA 2460496

A method of detecting a conformational change in a nucleic acid sample by treatment with oxidising agent for example KmnO4 to determine difference in formation or rate of formation of reaction product MnO2 or consumption or rate of consumption of KmnO4, which indicates conformational change in the test sample, in particular when the DNA is exposed to an environmental salt concentration and temperature or electrical current. The method investigates conformational change in the DNA, for example when DNA is exposed to an intercalating agent. The use of this method for investigating chemical compounds for conformational change in the DNA is also described. The conformational change is determined by colorimetric inorganic assay, by measuring absorbance by means of a spectrophotometer. The use of other oxidising agents besides permanganate comprising OsO4 or chromic acid or ozone gas, or peroxides or perbenzoic acids or electrical current, are described in the body of the specification.

La présente invention concerne un procédé permettant de détecter une modification de conformation affectant un échantillon d'acide nucléique. On a recours à un oxydant, essentiellement le KMnO¿4?, permettant de déterminer une différence, soit entre les formations ou les vitesses de formation du MnO¿2? obtenu, soit entre les consommations ou les vitesses de consommation du KMnO¿4?. Ces différences sont caractéristiques de modifications de conformation affectant l'échantillon test, plus particulièrement lorsque l'ADN est exposé à une atmosphère fortement saline et à la chaleur ou à un courant électrique. Le procédé permet de rechercher les modifications de conformation affectant l'ADN, notamment lorsque l'ADN est exposé à un agent intercalant. L'invention concerne également la mise en oeuvre du procédé pour la recherche de composés chimiques destinés aux modifications de conformation affectant l'ADN. Ces modifications de conformation sont attestées par un essai colorimétrique des inorganiques, par mesure de l'absorbance au spectrophotomètre. L'invention concerne aussi l'utilisation d'autres oxydants que le permanganate, et notamment l'OsO¿4?, l'acide chromique, l'ozone gazeux, les peroxydes, les acides perbenzoïques et le courant électrique.

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