@article{oai:repo.qst.go.jp:00082298, author = {Katsuya, Sato and Sanzen, Toshihiko and Yutaka, Ono and Narumi, Issay and Katsuya, Sato and Yutaka, Ono}, journal = {QST Takasaki Annual Report 2019}, month = {Mar}, note = {Deinococcus radiodurans is a representative strain of radioresistant bacteria and has extremely high resistance to various DNA damage caused by gamma rays, ultraviolet rays, desiccation, free radical-generating substances, and DNA cross-linkers. The previous studies revealed that the expression of a unique DNA repair-related protein, PprA, was up-regulated by a DNA damage response regulator, PprI, following DNA damage in D. radiodurans. However, the detailed functional site of PprI protein is poorly understood. In an effort to gain an insight into the role of PprI in DNA damage response mechanism in D. radiodurans, we generated luciferase reporter strains that were regulated by pprI expression plasmids. Firstly, we constructed a luciferase reporter DNA cassette consisted of an engineered firefly luciferase gene (FL) and an Escherichia coli spectinomycin resistance gene. Next, this luciferase reporter DNA cassette was integrated into the genome of the wild-type and the pprI-deleted mutant strains, and the resulting FL reporter strains were designated DARP and SIRP, respectively. Consequently, in strain DARP, FL reporter activity was enhanced following 2 kGy gamma irradiation at 2 h post-irradiation time. On the other hand, FL reporter activity in the pprI-deletion strain SIRP was not observed following irradiation. Like strain DARP, the FL reporter activity in the strain SIRP carrying the pprI expression plasmid was enhanced following irradiation by functional complementation with the pprI expression plasmid. This result suggests that the D. radiodurans PprI is a key protein for DNA damage response regulator.}, pages = {96--96}, title = {Construction of Luciferase Reporter Strains for Functional Analysis of DNA Damage Response Regulators}, volume = {QST-M-29}, year = {2021} }