Revista Colombiana de Biotecnología
versão impressa ISSN 0123-3475
VELASCO, Judith; ARAQUE², María e AYALA, Juan A.. Construction of mutants of Salmonella enterica for inactivation of invG/invE y ssaJ/ssaK genes of pathogenicity islands 1 and 2. Rev. colomb. biotecnol [online]. 2010, vol.12, n.2, pp. 55-66. ISSN 0123-3475.
To understand the genetic basis of Salmonella pathogenicity mechanisms, various methods have been described to manipulate and generate mutant genomic DNA with specific characteristics. In this study we report the construction of mutants from several serotypes of S. enterica, substitution and inactivation of genes invG/invE in SPI-1 gene and ssaJ/ssaK in SPI-2 by the technique of phage ? Red recombinase, as described by Datsenko and Wanner (2000). The gene deletion in SPI-1 and SPI-2 encodes proteins involved in the formation of type III secretion systems responsible for the invasion and intracellular survival of S. enterica in the host cells. The results of this work will allow in vivo studies to evaluate the possible attenuation of virulence of the mutant strains, as well as to provide new insights into the genetic mechanisms involved in the pathogenesis of diseases caused by these bacteria. Moreover, this technique is recommended to efficiently generate mutants of different serotypes of S. enterica in order to study the chromosomal genes and their products.
Palavras-chave : recombinasa red ?; mutación; islas de patogenicidad; Salmonella enterica; ? red recombinasa; mutation, pathogenicity islands; Salmonella enterica.