Sinorhizobium meliloti Optogenetics

Sinorhizobium meliloti is a gram-negative α-Proteus, which can fix atmospheric nitrogen during symbiosis with some host plants (such as alfalfa). S. meliloti and its pathogenic relatives are important models for studying host microbial interaction.

Microorganisms play an important role in shaping soil environment and promoting plant growth by interacting with plant roots. Due to the wide variety of cell types involved, coupled with dynamic and spatial heterogeneity, determining the causal contribution of specific factors remains elusive.

Creative BioMart developed a synthetic and light activated transcription control platform using blue light responsive DNA binding protein EL222 in nitrogen fixing soil bacterium alfalfa Rhizobium, and provided system construction services. By fine tuning the system, we have successfully realized the optical control of some material production routes.

How does it Works?

The LOV domain containing protein EL222 is a blue light sensing protein with a C-terminal helix turn helix (HTH) DNA binding domain representing LuxR type transcription regulators. El222 can act as an activator or repressor, This depends on whether its binding site is upstream (activator) or inside the promoter sequence (repressor). We constructed a blue light induced gene expression system in S. meliloti strain Rm8530. In the dark, EL222 is mainly a monomer and cannot bind to DNA. However, exposure to blue light will move the balance to dimer and enhance the binding and transcription with EL222 promoter.

Optogenetic Sinorhizobium meliloti System Construction

Service Content

Vector construction
Acquisition of tobacco aseptic seedlings
Protoplast isolation and polyethylene glycol-mediated transformation
Callus induction and adventitious bud differentiation of positive leaf disc
PCR detection of foreign gene in t0 generation transgenic seedlings

Service Description

Transgenic experiment

Receptor material varieties: we provide our own varieties and other varieties as receptor materials.
Type of receptor material: In order to realize the spatial and temporal control of production, we tried to implement light activated gene expression in S. meliloti strain Rm8530

Number of experiments

Vector construction: one vector was constructed for each foreign gene.
Gene Transformation: each transgenic vector was transformed into one receptor material.

Provide Results

Some pictures of genetic transformation and screening.
Differentiation and regeneration of adventitious buds.
Agarose gel electrophoresis of PCR gene detected by T0 generation.
Five or more lines were obtained in each transformation experiment.

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