New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria |
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Authors: | Arnaud Maryvonne Chastanet Arnaud Débarbouillé Michel |
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Affiliation: | Unité de Biologie des Bactéries Pathogènes à Gram Positif, Institut Pasteur, Paris, France. |
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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. |
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