Nicotiana tabacum Optogenetics
Comprehensive analysis of key plant cell processes requires command control of gene expression in time and space. Although some chemical induction systems showing satisfactory induction characteristics have been developed, they have inherent limitations in temporal and spatial resolution and may be related to toxic effects.
Creative BioMart describes here a synthetic light induction system for targeted control of gene expression in Nicotiana tabacum. The implementation of the system in the instantaneous determination of tobacco protoplasts leads to the strong induction of red light (660 nm) and can immediately return to the off state by subsequent far red light (740 nm). Using this characteristic similar to the toggle switch, the system can remain in the off state in the presence of 740 nm supplementary white light.
How it Works?
The synthetic switch is based on the red / far red light dependent interaction between phytochrome B (PhyB) and Arabidopsis phytochrome interaction factor 6 (PIF6), which is integrated into the transcription factors of light responsive division. On the one hand, PIF6 fuses with the DNA binding domain (BD), which binds its manipulation gene sites in the reporter construct upstream of the smallest promoter and reporter gene. On the other hand, PhyB is connected to the activation domain (AD) and nuclear localization sequence (NLS). Under the red light, PhyB-AD-NLS was recruited to PIF6 at the promoter site to switch the system to the on state. Only after the absorption of far red photons, the interaction between PhyB and PIF6 will terminate, resulting in the closure of gene expression.
Nicotiana tabacum Transgenic Service
| 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
- Rooting and strengthening of regenerated green plantlets
- PCR detection of foreign gene in t0 generation transgenic seedlings
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| Service Description |
Transgenic experiment
- Receptor material varieties: we provide our own varieties and other varieties as receptor materials.
- Type of receptor material: tobacco leaf.
Number of experiments
- Vector construction: one vector was constructed for each foreign gene.
- Gene Transformation: each transgenic vector was transformed into one receptor material, once.
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| 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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The Overview of the Biological Service Process
- Sign service contract
- The client transfers the target gene and makes payment
- We carried out transgenic experiments
- Deliver the experimental results to the customer, and the customer checks and accepts them
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