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Ramos Morales, Francisco Cardenal Muñoz, Elena Cordero Alba, Mar Baisón Olmo, Fernando 2025-02-03T07:32:57Z 2025-02-03T07:32:57Z 2015 Ramos-Morales F, Cardenal-Muñoz E, Cordero-Alba M, Baisón-Olmo F. Generation and use of site-directed chromosomal cyaA' translational fusions in Salmonella enterica. Methods Mol Biol. 2015;1225:93-104. doi: 10.1007/978-1-4939-1625-2_6. PMID: 25253250. https://hdl.handle.net/10630/37565 10.1007/978-1-4939-1625-2_6 CyaA from Bordetella pertussis is a calmodulin-dependent adenylate cyclase. Fusions to the catalytic domain of CyaA (CyaA’) are useful tools to detect translocation of type III secretion system effectors from Gram-negative pathogens like Salmonella enterica. These fusions are usually generated using plasmids with strong promoters. Here, we describe a protocol to insert the CyaA’-encoding sequence in a specific site in the bacterial chromosome in order to get a monocopy fusion whose expression is driven by the native promoter. We also describe the procedure to detect translocation of a CyaA’ fusion into mammalian cells. eng open access Translocación (Genética) Salmonella - Genética Bacterias gram negativas Bacterias - Metabolismo Generation and use of site-directed chromosomal cyaA’ translational fusions in Salmonella enterica. book part