Gene disruption in Lactobacillus plantarum strain 80 by site-specific recombination: isolation of a mutant strain deficient in conjugated bile salt hydrolase activity.
A chloramphenicol-resistance gene (cml) was introduced into the Lactobacillus plantarum gene encoding conjugated bile acid hydrolase (cbh) on a ColE1 replicon. This plasmid which is nonreplicative in Lactobacillus was used to transform L. plantarum strain 80. A homologous double cross-over recombination event resulted in replacement of the chromosomal cbh gene by the cml-containing cbh gene. The transformants obtained were unable to synthesize active conjugated bile acid hydrolase (Cbh). The Cbh- CmlR phenotype was stably maintained for more than 100 generations under nonselective conditions.
TNO Medical Biological Laboratory, Rijswijk, The Netherlands.
R J
RJ
Leer
H
H
Christiaens
W
W
Verstraete
L
L
Peters
M
M
Posno
P H
PH
Pouwels
D000581:Amidohydrolases / Q000096:biosynthesis / Q000235:genetics*; D002702:Chloramphenicol Resistance / Q000235:genetics; D005798:Genes, Bacterial; D007778:Lactobacillus / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011995:Recombination, Genetic; D015183:Restriction Mapping
D000581:Amidohydrolases; C023154:choloylglycine hydrolase