| RRC ID |
35379
|
| 著者 |
Biran A, Ben Yoav H, Yagur-Kroll S, Pedahzur R, Buchinger S, Shacham-Diamand Y, Reifferscheid G, Belkin S.
|
| タイトル |
Microbial genotoxicity bioreporters based on sulA activation.
|
| ジャーナル |
Anal Bioanal Chem
|
| Abstract |
A bacterial genotoxicity reporter strain was constructed in which the tightly controlled strong promoter of the Escherichia coli SOS response gene sulA was fused to the alkaline phosphatase-coding phoA reporter gene. The bioreporter responded in a dose-dependent manner to three model DNA-damaging agents-hydrogen peroxide, nalidixic acid (NA), and mitomycin C (MMC)-detected 30-60 min after exposure. Detection thresholds were 0.15 μM for MMC, 7.5 μM for nalidixic acid, and approximately 50 μM for hydrogen peroxide. A similar response to NA was observed when the bioreporter was integrated into a specially designed, portable electrochemical detection platform. Reporter sensitivity was further enhanced by single and double knockout mutations that enhanced cell membrane permeability (rfaE) and inhibited DNA damage repair mechanisms (umuD, uvrA). The rfaE mutants displayed a five- and tenfold increase in sensitivity to MMC and NA, respectively, while the uvrA mutation was advantageous in the detection of hydrogen peroxide. A similar sensitivity was displayed by the double rfaE/uvrA mutant when challenged with the pre-genotoxic agents 2-amino-3-methylimidazo[4,5-f]quinoline and 2-aminoanthracene following metabolic activation with an S9 mammalian liver fraction.
|
| 巻・号 |
400(9)
|
| ページ |
3013-24
|
| 公開日 |
2011-7-1
|
| DOI |
10.1007/s00216-011-5007-2
|
| PMID |
21533638
|
| MeSH |
Alkaline Phosphatase / genetics
Alkaline Phosphatase / metabolism
Animals
Biosensing Techniques / methods*
Electrochemical Techniques / methods
Escherichia coli / drug effects*
Escherichia coli / genetics*
Escherichia coli Proteins / genetics*
Escherichia coli Proteins / metabolism
Genes, Reporter
Hydrogen Peroxide / toxicity
Mitomycin / toxicity
Mutagenicity Tests / methods*
Mutagens / toxicity*
Nalidixic Acid / toxicity
Promoter Regions, Genetic
SOS Response, Genetics
|
| IF |
3.637
|
| 引用数 |
23
|
|
WOS 分野
|
BIOCHEMICAL RESEARCH METHODS
CHEMISTRY, ANALYTICAL
|
| リソース情報 |
| 原核生物(大腸菌) |
JW0374-KC
JW1172-KC
JW4019-KC
JW3024-KC |
