Serratia Marcescens Case Study

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Inducing Prodigiosin Transposon mutagenesis in Serratia Marcescens

Serratia Marcescens is an opportunistic pathogen, mainly of healthcare facilities but can also be found in many diverse environments. Serratia is a gram negative bacteria which can give it innate resistance to certain antibiotics, especially those that target peptidoglycan cell wall synthesis, due to its outer membrane. In an environment with different microorganisms competing for food Serratia holds a component that gives it another selective advantage. The bacteria contains a red pigment called prodigiosin, that has antibacterial, antifungal, and even antiprotozoal activity.
The pigment is produced due to quorum sensing of bacteria, when an appropriate level of N-hex
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pRL 27 plasmid map. tetAP: Plasmid promoter tnp: tn5 transposase element oriR6K: Origin of replication- allows replication in new bacterial cell aph: Kanamycin resistance oriT: Origin of transfer- allows transfer of plasmid elements into new cell

The E. coli WM2672 plasmid also contains genes for kanamycin resistance. This was used as a selective marker because if the Serratia accepted the plasmid then it will also have kanamycin resistance. The plain LB plates were used as an experimental control; The wild type, with prodigiosin production/no plasmid. The bacteria was plated in mid-log phase, this was done for two possible reasons. One being conjugation is highly efficient and successful during mid log phase and because kanamycin is an antibiotic that inhibits protein synthesis in growing bacteria by binding to the 30S subunit of the bacteria ribosome. This blocks the tRNA binding which stops the bacteria from making proteins for growth (Bacteriostatic). If the conjugation was successful the growing bacteria would be able to block kanamycin

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