Optogenetic-Lac E.coli system
The use of IPTG to control lac operons has been used to regulate gene expression in E. coli for numerous applications, including metabolic engineering and recombinant protein production. However, photogenetics provides unique capabilities that chemical inducers are difficult to obtain, such as ease of regulation, reversibility, dynamic induction intensity and spatial control.
Presently, Creative BioMart has not only established a complete R & D and design platform, but also established an experienced technical team. We are now able to build a light controlled lac operon regulation circuit for our customers. Blue light was used instead of IPTG to control the engineering metabolic pathway of chemical production in laboratory scale bioreactor.
Available Services at Creative BioMart
This is a simple particle system for light regulating gene expression. We placed lacI under the promoter of pDawn plasmid, so that the genes of interest were inhibited under light and induced in the dark. The circuit aims to induce gene expression in the dark to avoid the potential challenge of limited light penetration in high cell density fermentation.
| Phase |
Description |
Timeline |
| Subcloning into Expression Vector |
- The gene of interest (GOI) was amplified / isolated from the customer provided construct and subcloned into a suitable transfer vector.
- Insert sequence confirmation.
- Small induction of overexpression of target protein.
- Detection of recombinant proteins by SDS-PAGE and Western blot (if required, the customer must provide appropriate antibodies against proteins of interest).
|
3-5 weeks |
| Expansion and Purification |
- One liter of bacterial culture will be induced with IPTG and collected for protein purification.
- Ni-NTA used beads to purify GST labeled proteins with 6x his labeling or glutathione under natural or denatured conditions.
|
2 weeks |
| Deliverable |
- Transfer vector
- Recombinant Protein
- Documentation
- Culture with required yield
|
System Applications
- For recombinant protein production
- Induction of strains commonly used in metabolic engineering
- Study on bacterial movement and clustering
- Induction of protein degradation
- Biofilm and biomaterial formation
Blue Light VS IPTG
- The precise regulation of optogenetic system may be helpful to optimize the metabolic pathway of balanced chemical production.
- The circuit can regulate and control the gene expression level during fermentation, which can greatly reduce the number of constructs required for discovery.
- The system can be combined with future circuits for metabolic engineering, which responds perpendicular to light of different wavelengths
- The fine tuning and reversibility of gene expression provided by light may improve the yield of toxic proteins by adjusting its expression level, or improve the quality and solubility of proteins that are difficult to produce by adjusting its expression rate.
- In industrial fermentation, the elimination of IPTG (or other chemical inducers) will reduce the cost and increase the yield.
Creative BioMart is always devote us to provide high-quality and satisfactory service to our customers, if you are interested in our services or have some question, please feel free to contact us or make online inquiry.
