TY - JOUR
T1 - Bacterial bioluminescent emission from recombinant Escherichia coli harboring a recA::luxCDABE fusion
AU - Gu, Man Bock
AU - Min, Jiho
AU - Larossa, Robert A.
N1 - Funding Information:
This work was supported by the Korea Ministry of Education through Korea Research Foundation (grant # 97-F-20) and supported in part by the Korea Science and Engineering Foundation (KOSEF) through ADEMRC at Kwangju Institute of Science and Technology (K-JIST). The authors are grateful for their support.
PY - 2000/8/10
Y1 - 2000/8/10
N2 - This paper describes the quantitative evaluation of a bioluminescence assay for DNA damaging agents with respect to the linearity, sensitivity, specificity and dependence on the cell culture status. A recombinant bacterium, DPD2794, harboring a plasmid with a recA promoter fused to the luxCDABE operon, showed a very sensitive response to DNA-damaging stress. DPD2794 was found to show no noticeable response to non-mutagenic agents, i.e. phenol, except for some false responses appearing soon after injection. DPD2794 also showed a highly sensitive response to Mitomycin C, which was found to be a growth-stage-dependent response, not a growth-rate-dependent response. In addition, the relationship between the bioluminescence emitted in vivo, luciferase activity measured in vitro, and the amount of Lux proteins expressed was determined. The intensity of the bioluminescence emitted was found to be proportional to the luciferase activity in vitro, while the bioluminescence also seems to be correlated with the level of Lux proteins expressed in these Escherichia coli cells, up to 230 min post induction. Copyright (C) 2000 Elsevier Science B.V.
AB - This paper describes the quantitative evaluation of a bioluminescence assay for DNA damaging agents with respect to the linearity, sensitivity, specificity and dependence on the cell culture status. A recombinant bacterium, DPD2794, harboring a plasmid with a recA promoter fused to the luxCDABE operon, showed a very sensitive response to DNA-damaging stress. DPD2794 was found to show no noticeable response to non-mutagenic agents, i.e. phenol, except for some false responses appearing soon after injection. DPD2794 also showed a highly sensitive response to Mitomycin C, which was found to be a growth-stage-dependent response, not a growth-rate-dependent response. In addition, the relationship between the bioluminescence emitted in vivo, luciferase activity measured in vitro, and the amount of Lux proteins expressed was determined. The intensity of the bioluminescence emitted was found to be proportional to the luciferase activity in vitro, while the bioluminescence also seems to be correlated with the level of Lux proteins expressed in these Escherichia coli cells, up to 230 min post induction. Copyright (C) 2000 Elsevier Science B.V.
KW - DNA damaging agents
KW - Luciferase activity
KW - SOS response
KW - recA promoter
UR - http://www.scopus.com/inward/record.url?scp=0034632960&partnerID=8YFLogxK
U2 - 10.1016/S0165-022X(00)00100-7
DO - 10.1016/S0165-022X(00)00100-7
M3 - Article
C2 - 10899389
AN - SCOPUS:0034632960
VL - 45
SP - 45
EP - 56
JO - Journal of Proteomics
JF - Journal of Proteomics
SN - 1874-3919
IS - 1
ER -