Cloning And Transformation Of Pseudonomas aeruginosa Azoreductase Gene

Abstract

Pseudomonas aeroginosa (PA) is a gram negative aerobic rod that can cause serious, possibly life threatening, disease in humans, plants, and animals. In order to develop new and better treatments against PA, it is important to understand how this bacterium works to survive. Azoreductase from PA has quinone activity that aids the survival of PA, which can lead to an increase in antibiotic resistance. Using molecular biology approaches, we were able to successfully PCR amplify and clone the AzoR gene into a small, round strand of DNA called an expression plasmid vector (pET15b), transfer that plasmid into E.coli strain BL21, and then confirm that the E.coli were successfully transformed with the plasmid containing our gene of interest.