Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 45-56 |
| Number of pages | 12 |
| Journal | Journal of Biochemical and Biophysical Methods |
| Volume | 45 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2000 Aug 10 |
| Externally published | Yes |
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Keywords
- DNA damaging agents
- Luciferase activity
- recA promoter
- SOS response
ASJC Scopus subject areas
- Biochemistry
- Biophysics
Cite this
Bacterial bioluminescent emission from recombinant Escherichia coli harboring a recA : :luxCDABE fusion. / Gu, Man Bock; Min, Jiho; Larossa, Robert A.
In: Journal of Biochemical and Biophysical Methods, Vol. 45, No. 1, 10.08.2000, p. 45-56.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Bacterial bioluminescent emission from recombinant Escherichia coli harboring a recA
T2 - :luxCDABE fusion
AU - Gu, Man Bock
AU - Min, Jiho
AU - Larossa, Robert A.
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 - recA promoter
KW - SOS response
UR - http://www.scopus.com/inward/record.url?scp=0034632960&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034632960&partnerID=8YFLogxK
U2 - 10.1016/S0165-022X(00)00100-7
DO - 10.1016/S0165-022X(00)00100-7
M3 - Article
VL - 45
SP - 45
EP - 56
JO - Journal of Proteomics
JF - Journal of Proteomics
SN - 1874-3919
IS - 1
ER -