Investigation Of The Primary Transcriptome Of The Production Organism Pseudomonas Putida

Introduction: Pseudomonas putida is a nonpathogenic, Gram-negative bacterium and an excellent model organism for biotechnological applications. Due to its metabolic versatility, P. putida can grow in different environments including in extreme conditions. It has several genes to degrade xenobiotic compounds, a capability that renders the bacterium useful in bioremediation.

Finally, P. putida shows a high potential as a cell factory for the production of several compounds. Methods: Here, a differential RNA-sequencing approach (dRNA-seq) is used to gain new insights into the organization of the P. putida KT2440 transcriptome, in the presence of citrate or glucose as sole carbon source.

Results: A total of 7937 putative transcription start sites (TSSs) have been identified. 5’ RACE experiments have been performed to confirm putative TSSs, and 5’ UTR regions have been investigated for conservative motifs in promoter sequence. Finally, RNA regulatory elements have been identified including putative sRNAs and riboswitches.

Selected candidate riboswitch sequences have been tested for the study of the ligand-dependent regulatory mechanisms. Conclusions: By determination of the 5’-ends of transcripts, our study has allowed for the investigation of several biological features of P. putida.