Light-Inducible Recombinases System

The use of inducible recombinase can make specific cells in specific cell populations become recombination targets in response to the spatial pattern of light without changing the medium or waiting for the diffusion of chemical inducers. Light is a cheap inducer, which provides a powerful method for the precise spatio-temporal control of DNA modification.

Creative BioMart has developed a recombinase system in response to blue light activation based on E. coli. By dividing the recombinase gene into N-terminal and C-terminal fragments and connecting the sequence of photosensitive light dimer to each fragment, the recombinant enzyme can be sensitive to light. Under the light induction, the conformation of the light dimer changes, which makes it dimerize and binds the two fragments together. This well characterized recombinant enzyme tailored for E. coli is expected to become a powerful new tool for bacterial photogenetics.

Construction of the Light Inducible Recombinase System

At present, we have successfully constructed the framework of light induced Cre and Flp recombination system.

In order to make Cre sensitive to light, we divided it into N-terminal (nCre) and C-terminal (cCre) segments, and connected the light dimer to the inner end of each segment. When exposed to blue light, light dimerization changes the conformation to allow dimerization, thereby bringing CRE fragments together.

Flp is a tyrosine recombinase originally from Saccharomyces cerevisiae. The dimer used here is a designed heterodimer with a separate dimer interface.

Technology Application

The population dynamics was studied by controlling the spatial development of subspecies and establishing the interface between adjacent cells.
Changing the genetic state in the production of biomolecules in the application of metabolic engineering.
Cre recombinase can activate or inactivate multiple genes using structures similar to double fluorescence analysis.

Develop the Customized System

We provide bacterial expression system services, including codon optimization, gene synthesis, protein expression and two-step purification.

Service Name	Service Description
Gene synthesis and cloning, plasmid preparation	Codon optimization, gene synthesis, subcloning to expression vector Plasmid preparation and DNA QC
Protein expression test	The plasmid was transformed into a suitable bacterial expression strain Protein expression evaluation and optimization
Light induced protein expression and autolysis	Photoinduced expression in E.coli Autolysis of Escherichia coli cells in conical flask Label removal (if required)
QC & Delivery	Purity analysis and Detection( SDS-PAGE and Western Blot-tagged protein only) Protein delivery
Scale up and protein purification	Amplification Culture Preparation of crude extract Affinity purification Cut off fusion tag and remove protease (if necessary)

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