Scanning force microscopy studies of X-ray-induced double-strand breaks in plasmid DNA.

Abstract

We present an analysis of X-ray-induced damage in PhiX174 plasmid DNA, applying doses between D = 250 and 1,500 Gy. To analyse this damage in detail, the distribution of plasmid fragments after irradiation have been determined by scanning force microscopy. The results show that even for the lowest dose of D = 250 Gy, a significant amount of double-strand breaks are observed. For increasing dose, the percentage of small fragments increases and is accompanied by a shortening of the average fragment length from < L > = 1,400 nm for a dose of D = 250 Gy to < L > = 1,080 nm after irradiation with D = 1,500 Gy. The most crucial parameter, the average number of double-strand breaks per broken plasmid (<DSB(b)> ) has been determined for the first time for the applied doses. The results show that the average number of DSBs per broken plasmid <DSB(b)> increases almost linearly from a value of <DSB(b)> = 1.3 after irradiation with D = 250 Gy to <DSB(b)> = 1.7 after exposure to D = 1,500 Gy. The presented results show that the amount of DSBs induced by X-ray radiation in plasmid DNA can be calculated with high accuracy by means of scanning force microscopy, providing relevant information regarding the interaction of X-rays with DNA molecules.