New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria
M Arnaud, A Chastanet… - Applied and …, 2004 - Am Soc Microbiol
M Arnaud, A Chastanet, M Débarbouillé
Applied and environmental microbiology, 2004•Am Soc MicrobiolABSTRACT A shuttle vector designated pMAD was constructed for quickly generating gene
inactivation mutants in naturally nontransformable gram-positive bacteria. This vector allows,
on X-Gal (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside) plates, a quick colorimetric
blue-white discrimination of bacteria which have lost the plasmid, greatly facilitating clone
identification during mutagenesis. The plasmid was used in Staphylococcus aureus, Listeria
monocytogenes, and Bacillus cereus to efficiently construct mutants with or without an …
inactivation mutants in naturally nontransformable gram-positive bacteria. This vector allows,
on X-Gal (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside) plates, a quick colorimetric
blue-white discrimination of bacteria which have lost the plasmid, greatly facilitating clone
identification during mutagenesis. The plasmid was used in Staphylococcus aureus, Listeria
monocytogenes, and Bacillus cereus to efficiently construct mutants with or without an …
Abstract
A shuttle vector designated pMAD was constructed for quickly generating gene inactivation mutants in naturally nontransformable gram-positive bacteria. This vector allows, on X-Gal (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside) plates, a quick colorimetric blue-white discrimination of bacteria which have lost the plasmid, greatly facilitating clone identification during mutagenesis. The plasmid was used in Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus to efficiently construct mutants with or without an associated antibiotic resistance gene.
American Society for Microbiology