title: Chromosomal integration of the green fluorescent protein gene in lactic acid bacteria and the survival of marked strains in human gut simulations.
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.
authors.affiliation: Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen, UK. kps@rri.sari.ac.uk
authors.forename: K P
authors.initials: KP
authors.lastname: Scott
authors.forename: D K
authors.initials: DK
authors.lastname: Mercer
authors.forename: A J
authors.initials: AJ
authors.lastname: Richardson
authors.forename: C M
authors.initials: CM
authors.lastname: Melville
authors.forename: L A
authors.initials: LA
authors.lastname: Glover
authors.forename: H J
authors.initials: HJ
authors.lastname: Flint
mesh_terms: D001419:Bacteria / Q000254:growth & development; D002876:Chromosomes, Bacterial / Q000235:genetics; D015169:Colony Count, Microbial; D004251:DNA Transposable Elements; D004064:Digestive System / Q000382:microbiology*; D013293:Enterococcus faecalis / Q000235:genetics / Q000254:growth & development; D005243:Feces / Q000382:microbiology; D005285:Fermentation; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000235:genetics* / Q000254:growth & development*; D049452:Green Fluorescent Proteins; D006801:Humans; D019344:Lactic Acid / Q000378:metabolism; D013294:Lactococcus lactis / Q000235:genetics / Q000254:growth & development; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011995:Recombination, Genetic; D013291:Streptococcus / Q000235:genetics; D013753:Tetracycline Resistance / Q000235:genetics; D014169:Transformation, Bacterial
chemical_list: D004251:DNA Transposable Elements; D005819:Genetic Markers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D019344:Lactic Acid