At Sentebiolab, we provide state-of-the-art in vitro translation services for your research needs eliminating the use of live bacteria. In-vitro translation is the expression of the target protein or proteins in a cell-free environment which has certain advantages and uses among molecular biology applications. In-vitro translation applications can be used in rapid identification of proteins, folding studies, production of proteins that are toxic to the host cells and that cannot be expressed via recombinant expression methods, incorporation of modified or unnatural amino acids into the growing peptide chain and production of proteins that are prone to form inclusion bodies or proteolytic degradation.
In principle, in-vitro translation using cell-free extracts can be prepared from any kind of cells and give rise to the production of proteins from any mRNA molecule. In practice, cell-free extracts have only been developed from cells which have a high protein synthesis rate including rabbit reticulocytes or E.coli for eukaryotic and prokaryotic proteins, respectively.
E.coli is the most favored organism for large-scale protein expression and purification. However, some proteins cannot be expressed in E.coli as they are toxic to the organism. Furthermore, in order to express a protein in E.coli, the open reading frame coding the protein needs to be cloned into a suitable vector which is optimized for procaryotic protein expression. The plasmid then needs to be transformed into a suitable E.coli strain. These steps normally take about 3 days and the procedures are very labor intensive.
An alternative to this strategy is the utilization of in-vitro translation. Bioneer’s ExiProgen is a fully automated system that can efficiently perform in-vitro translation of 16 different proteins simultaneously. Each protein is expressed with a HisTag and purified in a scale of up to 100 µg using a standard kit. The system is also suitable for up to 500 µg/protein expression and purification using its maxi kit. Alternatively, up to 9 disulfide bond-bearing proteins can also be expressed and purified by this system.