Kinetics of the persistence of chromosomal DNA from genetically engineered Escherichia coli introduced into soil
Recorbet, G | Picard, C | Normand, P | Simonet, P | Unité de recherche Plantes et Systèmes de Culture Horticoles (PSH) ; Institut National de la Recherche Agronomique (INRA) | Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM) ; Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL) ; Université de Lyon-Université de Lyon-Ecole Nationale Vétérinaire de Lyon (ENVL)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)
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Показать больше [+] Меньше [-]Английский. Investigations to quantify bacterial survival and DNA persistence of a genetically engineered population of Escherichia coli introduced into soil microcosms were carried out. The survival of E. coli was monitored by plate counting and immunofluorescence methods, whereas the persistence of the DNA was evaluated by using a most-probable-number-polymerase chain reaction method. Whereas the E. coli population density declined below the plate-counting-technique detection threshold (10(2) CFU.g-1) after 15 days, 10(3) extracellular and 5 x 10(5) total DNA target sequences were still detected after 40 days. Additionally, the E. coli cell counts fell below the detection limit of the immunofluorescence method (10(5) cells.g-1) before the end of the experiment. Colony hybridizations did not reveal gene transfer to the indigenous microflora. These results confirm the persistence of residual E. coli target sequences that could not be detected by the classical cell counting method and offer promising applications for the environmental detection of microorganisms, either engineered, pathogenic, or released for beneficial effects.
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