Isolation and prevalidation of an Escherichia coli tester strain for the use in mechanistic and metabolic studies of genotoxins |
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| Institution: | 1. Department of Genetics, Faculty of Medical Sciences, UNL, R. de Junqueira 96, P-1300 Lisbon, Portugal;2. Faculty of Sciences and Technology, UNL, Monte de Caparica, Portugal;1. Department of Botany, Institute of Science, Banaras Hindu University, Varanasi, Uttar Pradesh, 221005, India;2. Virus Research and Diagnostic Laboratory, Government Institute of Medical Sciences, Greater Noida, Uttar Pradesh-9, 201310, India;1. Fujian Provincial Key Laboratory of Cancer Metastasis Chemoprevention and Chemotherapy, College of Chemistry, Fuzhou University, Fuzhou 350116, China;1. Lehrstuhl für Funktionelle Materialien, Physik-Department, Technische Universität München, James-Franck-Str. 1, 85748 Garching, Germany;2. Ningbo Institute of Materials Technology and Engineering, Chinese Academy of Sciences, 1219 Zhongguan West Rd, Ningbo 315201, PR China;3. Deutsches Elektronen-Synchrotron DESY, Notkestr. 85, 22603 Hamburg, Germany;4. Department of Fibre and Polymer Technology, KTH Royal Institute of Technology, Teknikringen 56-58, SE-100 44 Stockholm, Sweden;5. Heinz Maier-Leibnitz Zentrum (MLZ), Technische Universität München, Lichtenbergstr. 1, 85748 Garching, Germany |
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| Abstract: | We have isolated an Escherichia coli tester strain for the use in mechanistic and metabolic studies of genotoxins. We started with one of the more used and better characterized E. coli K-12 laboratory strains, AB1157. We isolated a lipopolysaccharide defective mutant of strain AB1886 which is an excision repair deficient derivative of AB1157 and introduced a newly constructed plasmid pKR11, encoding mucAB, resulting in strain MR2101/pKR11. A genotoxicity assay was designed, monitoring the reversion to arginine prototrophy and a preliminary validation was carried out against Ames tester strain TA100 with a set of diagnostic compounds. The results seem to indicate that strain MR2101/pKR11 is an adequate tester strain which can be a useful tool in mechanistic studies. Moreover, this strain can serve as mother strain to isolate improved and more especialized tester strains. |
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