TY - JOUR

T1 - Influence of plasma-generated reactive species on the plasmid DNA structure and plasmid-mediated transformation of Escherichia coli cells

AU - Lee, Geon Joon

AU - Choi, Min Ah

AU - Kim, Daewook

AU - Kim, Jun Young

AU - Ghimire, Bhagirath

AU - Choi, Eun Ha

AU - Kim, Seong Hwan

PY - 2017/9/11

Y1 - 2017/9/11

N2 - The influence of plasma-generated reactive species on the conformation of plasmid DNA (pDNA)and the transformation efficiency of Escherichia coli cells were studied. An atmospheric-pressureplasma jet (APPJ) was used to generate reactive oxygen and nitrogen species (RONS) in an aqueous solution. When E. coli cells were transformed, the transformation efficiency of E. coli with theAPPJ-treated plasmid was lower than with the APPJ-untreated plasmid. Transformation efficiencywas reduced due to structural modification and degradation of the pDNA by the APPJ. Plasmatreatment caused structural modification of the plasmid from the supercoiled form to the linearform, and also decreased the amount of plasmid by degrading the deoxyribonucleic acid (DNA)structure accompanied by disruption of nucleobases and DNA strand breakage. The formation oflinear plasmid from supercoiled plasmid by the APPJ treatment was verified through electrophoretic analysis of the NdeI restriction enzyme-cut supercoiled plasmid. The structural modificationand/or decrease in the amount of pDNA are attributed to the RONS from the plasma itself and tothose derived from the interaction of plasma radicals with the aqueous solution. The effect ofplasma treatment on the transformation efficiency of E. coli cells was more pronounced with thelinear plasmid than with the supercoiled plasmid, indicating that the linear plasmid is more vulnerable to RONS. Overall, these results revealed that plasma-generated RONS can modify the structuraland optical properties of bacterial pDNA, thus affecting its biological function.

AB - The influence of plasma-generated reactive species on the conformation of plasmid DNA (pDNA)and the transformation efficiency of Escherichia coli cells were studied. An atmospheric-pressureplasma jet (APPJ) was used to generate reactive oxygen and nitrogen species (RONS) in an aqueous solution. When E. coli cells were transformed, the transformation efficiency of E. coli with theAPPJ-treated plasmid was lower than with the APPJ-untreated plasmid. Transformation efficiencywas reduced due to structural modification and degradation of the pDNA by the APPJ. Plasmatreatment caused structural modification of the plasmid from the supercoiled form to the linearform, and also decreased the amount of plasmid by degrading the deoxyribonucleic acid (DNA)structure accompanied by disruption of nucleobases and DNA strand breakage. The formation oflinear plasmid from supercoiled plasmid by the APPJ treatment was verified through electrophoretic analysis of the NdeI restriction enzyme-cut supercoiled plasmid. The structural modificationand/or decrease in the amount of pDNA are attributed to the RONS from the plasma itself and tothose derived from the interaction of plasma radicals with the aqueous solution. The effect ofplasma treatment on the transformation efficiency of E. coli cells was more pronounced with thelinear plasmid than with the supercoiled plasmid, indicating that the linear plasmid is more vulnerable to RONS. Overall, these results revealed that plasma-generated RONS can modify the structuraland optical properties of bacterial pDNA, thus affecting its biological function.

U2 - 10.1063/1.4991081

DO - 10.1063/1.4991081

M3 - Journal article

VL - 122

JO - Journal of Applied Physics

JF - Journal of Applied Physics

SN - 0021-8979

M1 - 103303

ER -