Cell free protein synthesis (CFPS) has become a powerful technology platform, which can help to meet the current demand for protein expression. Saccharomyces cerevisiae is suitable for CFPS. It is a microorganism like E. coli and it can grow rapidly and cheaply in bioreactors or shakers under precise conditions. In addition, due to its eukaryotic properties, yeast is suitable for folding eukaryotic proteins and has previously shown the ability to modify after translation in vitro, such as glycosylation. As an expert in the field of cell-free expression, Creative BioMart has developed a yeast CFPS platform with high efficiency and high yield.
Click Here to See How to Select An Appropriate Cell-Free System→
This yeast CFPS is based on the Saccharomyces cerevisiae, a commonly used host cell for recombinant protein expression.
We constructed a plasmid regulated by T7 promoter, which can be used as a template for yeast CFPS after being inserted into E. coli. The expression system is a transcription/translation coupled system. The plasmid that contains the template sequence will be constructed in vitro before it can be added into yeast extract to complete translation and synthesize the target protein. At present, the reaction system we can provide is 15μ L. About 380ng/ml protein could be produced after a 2 h reaction.
In order to further save time and money for our customers, we optimize the processing steps. We evaluated the advantages of other processing steps, especially the treatment of MNase and in vitro transcription alone, since both steps involve expensive reagents. We found that in addition to the expected digestion of endogenous mRNA, MNase also digested ribosomal RNA, which is an undesirable result. Therefore, we chose to delete MNase pretreatment of yeast CFPS reaction. In addition, the Omega sequence of tobacco mosaic virus (TMV) was used to initiate translation, which coupled the transcription and translation steps and eliminated the in vitro transcription step
| Comparison of Conventional Yeast CFPS and the Improved Yeast CFPS | ||
| STEP | Conventional Yeast CFPS | Creative BioMart's Yeast CFPS |
| 1 | Shake flask cell culture | Bioreactor cell culture |
| 2 | Glass beads cell lysis | Homogenize cell lysis |
| 3 | Size exclusion buffer exchange | Dialysis buffer exchange |
| 4 | MNase treatment | Cell-free protein synthesis |
| 5 | Cell-free protein synthesis | |
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.
Reference
