New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria

Maryvonne Arnaud (1) , Arnaud Chastanet (1) , Michel Debarbouille (1)

1 Evolution et Génomique Bactériennes

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.

Mots clés: construction; recombination; thuringiensis; cloning; gene; listeria-monocytogenes; insertional mutagenesis

Mots clés: dna; staphylococcus-aureus; bacillus-subtilis

Domaines: Sciences du Vivant [q-bio]

Maryvonne Arnaud, Arnaud Chastanet, Michel Debarbouille. New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria. Applied and Environmental Microbiology, 2004, 70 (11), pp.6887-6891. ⟨10.1128/AEM.70.11.6887-6891.2004⟩. ⟨hal-02681992⟩