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coli et le vecteur de clonage qui y a été inséré peuvent donc être facilement identifiées. Ces jonctions sont uniques et spécifiques de E. coli ?fli. Elles ont donc été utilisées pour l'élaboration d'amorces (fli5 F et R ainsi que Fli3 F et R), TaqMan S_fli 5 et S_fli3) spécifiques ,
Cette variabilité pourrait également être expliquée par l'inoculum utilisé lors de ces premiers essais (environ 6.10 7 UFC) Afin de pallier cette fragilité de la qPCR, il pourrait être envisagé d'augmenter la quantité de bactéries donneuses inoculées sans toutefois dépasser les doses (10 9 à 10 11 UFC par jour) utilisées pour l'administration de probiotiques (Wolvers et al. 2010) Une autre solution serait d'examiner les contenus intestinaux où les concentrations de la souche donneuse et d'éventuels transconjugants pourraient être plus élevées ,
une souche possédant un double marquage fluorescent permet de séparer les bactéries donneuses des transconjugants. La mise au point d'une telle souche a été possible lors de ce travail En effet, E. coli ?fli (pB10::rfp) porte un marquage chromosomique gfp et plasmidique rfp. L'analyse de cette souche en CMF n'a pas permis de détecter la fluorescence rouge liée à la RFP plasmidique. Ceci pourrait être expliqué par une trop faible expression de la RFP par les cellules bactériennes après 24 heures de croissance ,
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