Team:Freiburg/Project/pRIG15 15

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pRIG15_15

Salmonella Typhimurium is a pathogen causing gastrointestinal infections and which is particularly dangerous because it already gained resistence against multiple antibiotics. ¹⁾ In collaboration with the group from Prof. Dr. Hust from the University of Braunschweig we were able to get an expression vector containing the sequence for a putative dihydroxyacid dehydratase and an additional vector bearing the respective antibody, anti-putative dihydroxyacid dehydratase Bba_K1621007. ²⁾

To insert the sequence for the Salmonella dehydroxyacid dehydratase into pSB1C3 we designed Gibson primers with compatible overhangs that also included the start codon ATG. This fragment was amplified via PCR (Link zum Labjournal-Eintrag) from the expression vectors we got from Prof. Dr. Hust and then assembled with the digested pSB1C3 backbone using Gibson assembly. To prove correct insertion of our fragment we did a test digest (Link Labjournal) and sent the whole plasmid for sequencing

¹⁾ Little et al. 2008. Campylobacter and Salmonella in raw red meats in the United Kingdom: prevalence, characterization and antimicrobial resistance pattern, 2003-2005. Food microbiology

²⁾ Meyer et al. 2012. Identification of immunogenic proteins and generation of antibodies against Salmonella Typhimurium using phage display. BMC biotechnology