title: New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria.
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-beta-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.
authors.affiliation: Unité de Biologie des Bactéries Pathogènes à Gram Positif, Institut Pasteur, Paris, France.
authors.forename: Maryvonne
authors.initials: M
authors.lastname: Arnaud
authors.forename: Arnaud
authors.initials: A
authors.lastname: Chastanet
authors.forename: Michel
authors.initials: M
authors.lastname: Débarbouillé
mesh_terms: D000483:Alleles; D001482:Base Composition; D005822:Genetic Vectors; D006094:Gram-Positive Bacteria / Q000235:genetics*; D010957:Plasmids; D011995:Recombination, Genetic; D014169:Transformation, Bacterial