Chromosomal integration of the green fluorescent protein gene in lactic acid bacteria and the survival of marked strains in human gut simulations |
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Authors: | Scott K P Mercer D K Richardson A J Melville C M Glover L A Flint H J |
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Affiliation: | Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen, UK. kps@rri.sari.ac.uk |
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Abstract: | An integration vector was constructed to allow introduction of the gfp gene into the chromosomes of Gram-positive bacteria. Integration depends on homologous recombination between a short 458-nt sequence of the tet(M) gene in the vector and a copy of Tn916 in the host chromosome. Strains of Lactococcus lactis IL1403, Enterococcus faecalis JH2-SS, and Streptococcus gordonii DL1 stably marked with single chromosomal copies of the gfp were readily visualised by epifluorescence microscopy. The marked L. lactis strain survived poorly in a continuous culture system inoculated with human faecal flora, while the laboratory E. faecalis strain was lost at approximately the dilution rate of the fermenter. |
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Keywords: | gfp Chromosomal integration Gram-positive bacterium Tetracycline resistance Tn916 |
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