High Level Protein Expression in Cyanobacteria
The design of biological processes using cyanobacteria as host microorganisms is challenging, but considering the potential of Cyanobacteria to produce various valuable compounds, it is also attractive to culture with CO2 and simple nutrients due to their photosynthetic autotrophic characteristics.
Creative BioMart improved a green light regulated T7 RNA polymerase expression system and introduced it into Synechocytis sp. PCC6803, which provides a new and multifunctional high expression system for cyanobacteria, and realizes gene expression in Cyanobacteria with high expression level without using any chemical inducer.
How does it Works?
Under green light irradiation, the green light sensor CcaS activates the response regulator CcaR to induce the promoter cpcg2. T7 RNAP transcribes the T7 promoter to express the inserted gene. Under red light irradiation, CcaS inactivates CcaR, thereby inhibiting the expression of T7 RNAP and the expression of inserted genes under T7 promoter.
Our Service
| Service Name |
Service Description |
Deliverable |
| Preparation of the vector |
- Digestion with restriction enzyme, dephosphorylation.
- Gel purification
|
- Cloning vector
- QC data
- Final report
- Protein product( quantity as per customer requirements)
|
| Preparation of target DNA |
- Codon optimization
- Plasmid purification
- Gel purification
|
| Subclone construction |
- Subclone the target sequence to the pET vector
- Transformation of non-expression host bacteria
- Screening positive clones
- Clone sequencing to determine ORF
|
| Protein expression |
- Transformation of expression host E. coli strain that contains T7 RNA polymerase gene
- Plasmid stability test (optional)
- The cellular and subcellular components, expression time, temperature conditions, protein solubility and activity test
- Qualitative and quantitative analysis of protein products
|
| Scale up and protein purification |
- Amplification Culture
- Preparation of crude extract
- Affinity purification
- Cut off fusion tag and remove protease (if necessary)
|
How do we Ensure the Quality of the Expression System
- Codon optimization. We will carry out codon optimization, involving gene synthesis, vector construction, gene guaranteed transcription, mRNA translation, post-translational modification.
- Transcriptional analysis of green light regulated T7 RNAP expression was performed by quantitative reverse transcription PCR
The expression level of T7 RNAP system was evaluated under separate red light or red and green light irradiation.
Service Advantages
- The success rate of the project is greater than 95%.
- One stop technical service
- Codon optimization: comprehensively consider more than 20 factors in the process of gene synthesis, vector construction, gene transcription, mRNA translation and modification.
- Large scale preparation and fermentation: we have 10L, 100L, 500L and 1000L fermentation tanks, which can deliver gram grade high-purity target protein to customers in a short time.
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.
