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Use of the Golden Gate Cloning Technique for Plasmid Construction to Generate Conditional Lethal Mutants in C. difficile.

Methods in molecular biology (Clifton, N.J.) - 1 Jan 2026

Malet-Villemagne Jeanne, Zielinski Marie-Emeline, Parfut Assilina, Candela Thomas

Abstract excerpt

Conditional lethal mutants are a valuable tool for studying essential genes in bacteria, as they allow for the controlled expression of the gene of interest under specific conditions. In this study, we present a protocol for generating conditional lethal mutants in Clostridioides difficile using the Golden Gate cloning technique. The strategy involves two steps: (1) introducing an inducible copy of the gene of...

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PMID: 42096147
DOI: 10.1007/978-1-0716-5328-9_3

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