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Consistent biosynthesis of desired secondary metabolites (SMs) from pure microbial cultures is often unreliable. In a proof-of-principle study to induce SM gene expression and production, we describe mixed “co-culturing” conditions and monitoring of messages via quantitative real-time PCR (qPCR). Gene expression of model bacterial strains ( Pseudomonas aeruginosa  PAO1 and  Roseobacter denitrificans  Och114) was analyzed in pure solo and mixed cocultures to infer the effects of interspecies interactions on gene expression  in vitro , Two  P. aeruginosa  genes ( PhzH  coding for portions of the phenazine antibiotic pathway leading to pyocyanin (PCN) and the  RhdA  gene for thiosulfate: cyanide sulfurtransferase (Rhodanese)) and two  R. denitrificans  genes ( BetaLact  for metallo-beta-lactamase and the  DMSP  gene for dimethylpropiothetin dethiomethylase) were assessed for differential expression. Results showed that  R. denitrificans DMSP  and  BetaLact  gene expression became elevated in a mixed culture. In contrast,  P. aeruginosa  co-cultures with  R. denitrificans  or a third species did not increase target gene expression above control levels. This paper provides insight for better control of target SM gene expression  in vitro  and bypass complex genetic engineering manipulations.

Co-Cultures ofPseudomonas aeruginosaandRoseobacter denitrificansReveal Shifts in Gene Expression Levels Compared to Solo Cultures