Assay for reactive oxygen species-induced DNA damage: measurement of the formamido and thymine glycol lesions.
Biochim Biophys Acta
; 1454(1): 80-8, 1999 May 31.
Article
en En
| MEDLINE
| ID: mdl-10354517
A 32P-postlabeling assay has been developed for the simultaneous detection of the thymine glycol lesion and the formamido remnant of pyrimidine bases in DNA exposed to reactive oxygen species (ROS). The formamido lesion is a principal lesion produced in X-irradiated DNA oligomers when oxygen is available to mediate the damage process. Production of the well-known thymine glycol lesion is less dependent on the concentration of oxygen. These two lesions have the common property that they make the phosphoester bond 3' to the modified nucleoside resistant to hydrolysis by nuclease P1. Our assay uses 32P-postlabeling to measure these lesions in the form of modified dimers obtained from DNA by nuclease P1 digestion. Appropriate carriers and internal standards have been chemically synthesized to improve the reliability and accuracy of the assay. The measurements were accomplished on 1-microgram samples of DNA.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Nucleósidos de Pirimidina
/
Timina
/
Daño del ADN
/
Especies Reactivas de Oxígeno
Idioma:
En
Revista:
Biochim Biophys Acta
Año:
1999
Tipo del documento:
Article
País de afiliación:
Estados Unidos
Pais de publicación:
Países Bajos