Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 70, Issue 11, Pages 6887-6891Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.70.11.6887-6891.2004
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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.
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