Construction of Plasmid Vectors for Screening Replicons from Gram-Positive Bacteria and Their Use as Shuttle Cloning Vectors

Abdellah Benachour (1, 2) , Yanick Auffray (1, 2) , Axel Hartke (2)

1 U2RM - Unité de Recherche Risques Microbiens
2 LME - Laboratoire de Microbiologie de l’Environnement

Plasmids play a central role in engineering recombinant bacteria because they are the primary vehicles used to manipulate targeted sequences. In some cases, bacteria of interest are poorly provided with suitable tools for these molecular or genetic manipulations. In this context, we constructed from two shuttle cloning vectors, pUCB2871 and pUCB2872, the basic vectors pUCB30 and pUCB31, which could represent suitable tools to isolate replicons from Gram-positive bacteria. These plasmid vectors are characterized by the following after-features: (a) the pUC origin of replication is unable to replicate in Gram-positive bacteria; (b) an erythromycin-resistance encoding gene that is functional in both Gram-negative and -positive bacteria; (c) the pUC19 multiple cloning site (MCS) within the lacZα reporter gene; and (4) an additional multiple cloning site (MCS). Cloning replicons from Gram-positive bacteria in this additional MCS would allow the derivative vectors to function directly as shuttle cloning vectors.

Mots clés: Multiple Cloning Site; Rolling Circle Replication; Segregational Stability; Plasmid Replicon; Antibiotic Resistance Cassette

Domaines: Sciences du Vivant [q-bio]; Biochimie, Biologie Moléculaire

Abdellah Benachour, Yanick Auffray, Axel Hartke. Construction of Plasmid Vectors for Screening Replicons from Gram-Positive Bacteria and Their Use as Shuttle Cloning Vectors. Current Microbiology, 2007, 54 (5), pp.342-347. ⟨10.1007/s00284-006-0358-1⟩. ⟨hal-02180998⟩