Project Description

Project Description

Inducible promoter systems used in bacterial engineering are powerful tools for fine-tuned control of gene expression. Heightened inducibility and tighter transcriptional control allows for greater efficacy in a biomanufacturing context, granting larger yields of desired genetic products while minimizing toxic or interfering protein production. Current inducible promoter systems like the TetR-Ptet, exhibit high basal levels of expression, or “leakiness”, in an array of microbial chassis that negatively impact the quality of inducibility and repression of target genes. The TetR-Ptet system is functional in methanotrophic bacteria, like Methylococcus capsulatus, but other, less leaky inducible promoters would facilitate advanced metabolic engineering of these microbes for greenhouse gas mitigation applications. Our team is adapting the Jungle Express (JEx) inducible promoter system for use in M. capsulatus. The JEx system was previously shown to exhibit tighter transcriptional regulation and a 4-fold greater magnitude of inducibility compared to the TetR-Ptet system in E. coli, and utilizes a cheap and accessible inducing molecule, crystal violet. The flexibility and versatility of the JEx system lends itself to unprecedented control in genetic engineering of methanotrophic bacteria, which could enable their use for methane mitigation and carbon capture .


References


  • Jungle Express Paper
  • Ruegg TL, Pereira JH, Chen JC, DeGiovanni A, Novichkov P, Mutalik VK, Tomaleri GP, Singer SW, Hillson NJ, Simmons BA, Adams PD, Thelen MP. Jungle Express is a versatile repressor system for tight transcriptional control. Nat Commun. 2018 Sep 6;9(1):3617. doi: 10.1038/s41467-018-05857-3. Erratum in: Nat Commun. 2018 Oct 30;9(1):4596. doi: 10.1038/s41467-018-07111-2. PMID: 30190458; PMCID: PMC6127294.