Proton-induced direct and indirect damage of plasmid DNA
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Clustered DNA damage induced by 10, 20 and 30 MeV protons in pBR322 plasmid DNA was investigated. Besides determination of strand breaks, additional lesions were detected using base excision repair enzymes. The plasmid was irradiated in dry form, where indirect radiation effects were almost fully suppressed, and in water solution containing only minimal residual radical scavenger. Simultaneous irradiation of the plasmid DNA in the dry form and in the solution demonstrated the contribution of the indirect effect as prevalent. The damage composition slightly differed when comparing the results for liquid and dry samples. The obtained data were also subjected to analysis concerning different methodological approaches, particularly the influence of irradiation geometry, models used for calculation of strand break yields and interpretation of the strand breaks detected with the enzymes. It was shown that these parameters strongly affect the results.
KeywordsProton radiation DNA plasmid Direct and indirect effects Clustered damage Repair enzymes
This work has been supported by ESA Project No. ESTEC/ITT AO/1-7146/12/NL/GLC “Radiation Biological End Effects Models and Interfaces to Physics Models” and MEYS CR Project No. LD12008. L. Vyšín and L. Juha appreciate a partial financial support provided by the Czech Science Foundation under the Grant No. 13-28721S. M. Vlk is grateful for a partial financial support by MEYS CR Project No. LK21310. The irradiation has been realized at the CANAM (Center of Accelerators and Nuclear Analytical Methods) supported by the MEYS CR Project No. LM 2011019. The authors also appreciate valuable comments of Dr. Amitava Adhikary.
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