||Visualization of DNA Repair Process in Mammalian Cells Transformed by Plasmid DNA Exposed to X-rays in vitro
中上, 裕貴 ,
小畑, 結衣 ,
神長, 輝一横谷, 明徳
Radiation damage to DNA and its repair mechanism have been studied from both molecular (in vitro) and cellular (in vivo) levels. Although irradiation and chemical analysis of isolated DNA samples give us evidences of chemical aspects of damage types and their yields, it would be difficult to infer their repair-susceptibility. On the other hand, it is difficult to specify DNA damage types, except double strand breaks, in irradiated cells, and furthermore whole-cell irradiation inevitably causes damage not only in DNA but also organelle which may modify the DNA damage type. To highlight the cellular effects of DNA damage induced by various types of radiation in distinction from other damage in cytoplasm, we transformed X-ray irradiated plasmid DNA, which is constructed of a fluorescence protein gene, EGFP, into non-irradiated mammalian cells using lipofectamine. The repair kinetics of the irradiated plasmids in the cell could be visualized under microscopic imaging as recovery of fluorescence emission from the transformed cells. Using an agarose gel electrophoresis method, the yields of single- and double- strand breaks were also quantified, and compare the imaging data. We will show preliminary data obtained by this novel technique applied for plasmids exposed to X-rays, or treated with a nicking or restriction enzyme.
MICROS 2017 - 17th International Symposium on Microdosimetry 参加