Chromosomal integration of the green fluorescent protein gene in lactic acid bacteria and the survival of marked strains in human gut simulations

An integration vector was constructed to allow introduction of the afp gene into the chromosomes of Gram-positive bacteria. Integration depends on hom ologous recombination between a abort 458-nt sequence of the tet(M) gene in the vector and a copy of Tn916 in the host chromosome. Strains of Lactococ cus lactis IL1403, Enterococcus faecalis JH2-SS, and Streptococcus gordonii DL1 stably marked with single chromosomal copies of the gfp were readily v isualised by epifluorescence microscopy. The marked L. lactis strain surviv ed poorly in a continuous culture system inoculated with human faecal flora , while the laboratory E. faecalis strain was lost at approximately the dil ution rate of the fermenter. (C) 2000 Published by Elsevier Science B.V. Al l rights reserved.