Light Induced Autolysis System
E.Coli has been well characterized in molecular genetics, physiology and expression system, and is widely used to produce heterologous proteins. However, the traditional protein expression and purification process is complex and cumbersome, and these proteins are usually expressed as intracellular proteins in E. coli, which need to destroy the cell wall to obtain the target protein. In addition, it is usually necessary to add chemical inducers (such as IPTG) to achieve the maximum yield of the required protein. IPTG is expensive, toxic and difficult to remove during downstream operation. These problems of heterologous proteins seriously hinder the research and application of proteins.
In order to solve this problem for customers, Creative BioMart uses a new strategy of expressing target proteins using a cost-effective and simple program to meet the growing demand for protein production. We will combine the SRRz autolysis system and photoinduction system of autophagy, which provides a new idea for the efficient and economic production of protein.
Service Procedure at Creative BioMart
In the protein induction strategy, photogenetic methods have proved that light can accurately control cell function. The unique variability of spike lasers, including wavelength and intensity, allows specific triggering of cellular events in a non-invasive and high-resolution spatio-temporal manner. We inserted the SRRz gene of autophagy lambda downstream of the T7 promoter. Different from the traditional induced expression system, the T7 promoter is activated by light rather than IPTG.
| Service Name |
Service Description |
Deliverable |
| Gene synthesis and cloning, plasmid preparation |
- Codon optimization, gene synthesis, subcloning to expression vector
- Plasmid preparation and DNA QC
|
- Cloning vector
- QC data
- Final report
- Protein product (quantity as per customer requirements)
|
| 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
- Purification to achieve the required protein quantity and purity
- 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)
|
We provide construction services and protein expression services, including codon optimization, gene synthesis, protein expression and two-step purification. If you need other services, such as protein refolding, label removal and endotoxin removal services, you can also make additional choices. Through our proprietary codon optimization software and our experienced scientific team, we guarantee the delivery of high-quality final products.
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.
