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A Novel methodology for characterizing strand-break termini and damaged bases in plasmid DNA exposed to ionizing radiation

Akamatsu, Ken

We have developed a de novo methodology to characterize radiation damage in DNA. An enzyme system consisting of snake venom phosphodiesterase (SVPD) and calf intestine alkaline phosphatase (CIAP) was used to examine the 3'termini of strand break sites. In this study, we hypothesized that the strand-break termini can be divided into two categories. The apparent radiation-chemical yield (G-value) can be estimated using the level of intact 2'-deoxynucleotides produced during a 15-min incubation with SVPD. The G-value for total strand breaks in dried DNA irradiated with $$^{60}$$Co $$gamma$$-rays was estimated to be 0.1 micromol/J. Moreover, the G-values of CIAP-dependent and CIAP-independent SVPD sites were estimated to be 0.078 and 0.024 micromol/J, respectively. These values suggest that 3'-phosphate termini are more likely to be produced than 3'termini without phosphate.

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Category:Biochemical Research Methods

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