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Survival and transfer in the human gut of poorly mobilizable (pBR322) and of transferable plasmids from the same carrier E. coli
Authors:Bonnie Marshall  Susan Schluederberg  Chikanori Tachibana  Stuart B Levy
Institution:Departments of Molecular Biology and Microbiology and of Medicine, Tufts University School of Medicine, Boston, MA 02111 U.S.A.
Abstract:We examined the survival of a host Escherichia coli K-12 bacterium containing two transferable plasmids (pLM2, pSL222-4) and one poorly mobilizable plasmid (pBR322), and the transfer of these three plasmids to endogenous bacteria in the human intestinal tract. The survival of this plasmid-carrying host organism in four human volunteers was 3.5 to 6 days at recovery rates of 10?1 to 10?4. This finding was similar to our previous survival data on the same organism bearing a single plasmid. The K-12 strain appeared to be under a strong selective disadvantage in the human gut, since, even when bearing a tetracycline-resistant plasmid, its titer did not increase despite the administration of tetracycline. Studies of transferability showed that, while the transfer-depressed incFII plasmid pSL222-4 transferred at a frequency of 10?1 in culture, its transfer in the human gut was much less frequent. The number of new recipients per donor cell ingested was about 10?5, which included new recipients arising by multiplication. The recovery of pSL222-4 transcipients was enhanced by the administration of tetracycline on day 6. Neither the transfer-repressed, broad host range incP plasmid pLM2, nor the plasmid pBR322, could be detected in any endogenous host bacteria. Using the transfer and mobilization frequencies obtained in culture and the number of new recipients of pSL222-4 in the intestinal tract, we estimated that any in vivo mobilization of pBR322 to a new recipient could not occur at a frequency higher than 10?12.
Keywords:Human volunteers  pBR322  pLM2  pSL222-4 plasmids  cloning vectors  recombinant DNA safety  antibiotic resistance  amp  ampicillin  cm  chloramphenicol  kan  kanamycin  Mac  MacConkey  Md  megadalton  nal  nalidixic acid  PEA  phenylethylalcohol  rif  rifamycin  sm  streptomycin  sp  spectinomycin  sul  sulfanilamide  tc  tetracycline  []  indicates the plasmid-carrier state
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