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dc.contributor.authorMesa-Pereira, Beatriz-
dc.contributor.authorMedina, Carlos-
dc.contributor.authorCamacho, Eva María-
dc.contributor.authorFlores, Amando-
dc.contributor.authorSantero, Eduardo-
dc.identifierdoi: 10.1111/1751-7915.12153-
dc.identifiere-issn: 1751-7915-
dc.identifier.citationMicrobial Biotechnology 8(1): 169-176 (2015)-
dc.descriptionThis is an open access article under the terms of the Creative Commons Attribution License.-
dc.description.abstractIn order to increase the cytotoxic activity of a Salmonella strain carrying a salicylate-inducible expression system that controls cytosine deaminase production, we have modified both, the vector and the producer bacterium. First, the translation rates of the expression module containing the Escherichia colicodA gene cloned under the control of the Pm promoter have been improved by using the T7 phage gene 10 ribosome binding site sequence and replacing the original GUG start codon by AUG. Second, to increase the time span in which cytosine deaminase may be produced by the bacteria in the presence of 5-fluorocytosine, a 5-fluorouracyl resistant Salmonella strain has been constructed by deleting its upp gene sequence. This new Salmonella strain shows increased cytosine deaminase activity and, after infecting tumour cell cultures, increased cytotoxic and bystander effects under standard induction conditions. In addition, we have generated a purD mutation in the producer strain to control its intracellular proliferation by the presence of adenine and avoid the intrinsic Salmonella cell death induction. This strategy allows the analysis and comparison of the cytotoxic effects of cytosine deaminase produced by different Salmonella strains in tumour cell cultures. We have increased the cytotoxic activity of a Salmonella strain carrying a salicylate-inducible expression system that controls cytosine deaminase production. To that end, we have increased the cytosine deaminase translation rates and the time span in which it is produced by generating a 5-FU resistant Salmonella. The inclusion of a purD mutation in the producer strain has also allowed us to control its intracellular proliferation and analyse the effects of cytosine deaminase production by different strains in tumour cell cultures.-
dc.description.sponsorshipThis work was supported by the Grant ‘Proyecto de Excelencia P07-CVI02518’ from the Andalusian government and by a fellowship from the Andalusian government to B. M.-P. C. M. holds a JAE DOC contract from the Spanish National Research Council (CSIC).-
dc.publisherJohn Wiley & Sons-
dc.relation.isversionofPublisher's version-
dc.titleImproved cytotoxic effects of Salmonella-producing cytosine deaminase in tumour cells-
dc.description.versionPeer Reviewed-
dc.contributor.funderJunta de Andalucía-
dc.contributor.funderConsejo Superior de Investigaciones Científicas (España)-
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