In the past decades, cell-free expression has proved to be a promising platform for the synthesis of many different proteins, including membrane proteins, toxic proteins and proteins with post-translational modifications.
Insect cell-free expression system (ICE) is the most widely used eukaryotic system. ICE is better than rabbit reticulocyte lysate expression system (RRL) in terms of glycosylation. The advantage of this method is that the core glycosylation of ice does not need to add microsome membrane, and it has been proved that microsome membrane provides the necessary isolation and enzyme for proper post-translation modification in RRC.
Click Here To See The comparison of each cell-free protein synthesis systems→
Creative BioMart is a global pioneer and leader in the use of insect systems for the expression of cell-free membrane proteins. We optimized podophera frugiperda Sf21 cell line and used it as the source of extract. In addition, we constructed a vector template with 5'-UTR fragment of the baculovirus polyhedron gene. Through this system, we can express recombinant proteins, especially membrane proteins efficiently.
The extract is prepared from the podophera frugiperda Sf21 cell line by standard procedure. We mix all the ingredients needed for transcription and translation into ice extract.
| Extract Source | Template Form | Promoter | Productivity | Reaction Time |
| Sf21 cell line | DNA | T7 Promoter | Up to 7 mg/ml | 4 hours at 28-30°C |
When choosing our services, the client needs to mail or authorize us to synthesize DNA templates. The target sequence needs to be cloned downstream of the T7 promoter. We also developed a vector containing the 5 '- 3' untranslated (UTR) sequence of baculovirus polyhedron gene to improve translation efficiency (if you need to use our vector, please contact us for a plasmid map). After the template was added to the extract, the protein expression was up to 7 mg/ml after incubation at 28-30°C for about 4 hours.
| Transcription/translation-Coupled RRL Process | ||
| 1 | Add DNA template and perform coupled transcription and translation | 4 hours at 28-30°C |
| 2 | Separate translation products by SDS-PAGE | 1.5 hours |
| 3 | Fix and dry gel | 2 hours |
| 4 | Autoradiography | 3 hours, -70°C |
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.
