The most classic E. coli cell-free protein synthesis system is E. coli S30 extract system. The cell lysate used in the system is prepared by using the mutant E. coli B with deficient OmpT endoproteinase and lon protease activity. At Creative BioMart, this system has been implemented for many years. After years of practice, we have completed the optimization of the classic E. coli S30 extract system. Considering the different expression needs of our clients, we now provide expression services based on circular DNA and linear DNA templates.
This service allows successful transcription/translation of linear DNA templates in E. coli S30 cell lysate.
| Clients Provide | A. Linear DNA |
| B. Submit DNA sequence information online (We need your authorization to synthesize the sequence) | |
| C. DNA synthesized by PCR, linked oligonucleotides, RNA produced in vitro and prokaryotic RNA can be used as templates, but the final yield is at risk of decline |
*Client can choose to provide us with any form of the above templates as long as the sequence contains prokaryotic E. coli-like promoter such as lacUV5, tac, λPL (con) and λ-PR.
Identification of cloned or synthesized genes from linear or closed circular DNA.
Rapid gene structure mapping
Rapid verification of mutation in vitro.
Synthesis of truncated gene products for functional domain mapping and epitope mapping.
Expression of genes lacking suitable E.coli promoter in RNA produced in vitro
Client's plasmid DNA template will be transcription and translation via the E. coli Extract Protein Synthesis Service for Circular Templates. This system is more active than the one for linear DNA. Again, the provided plasmid or the corresponding sequence should contain prokaryotic E. coli-like promoters such as lacUV5, tac, λPL (con) and λ-PR.
The most common application of expressing circular DNA in E. coli lysate is to synthesize a small amount of radiolabeled proteins. In addition, the technology can also be used for the study of multiple functions of transcription and translation, as a tracer in protein purification, identifying cloned DNA gene products or locating peptides to specific DNA fragments, and screening compounds that inhibit bacterial translation.
In addition to the classic expression services above, we also provide a high yield T7 Regulated E. coli Extract Protein Synthesis Service. In the system, the template can be provided in the form of plasmid DNA, Liner DNA or mRNA.
Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.
