This gene synthesis step is very important because it allows us to optimize rare codons for E. coli expression system. It significantly increases production yields in many cases and especially with E. coli. This strategy has proven to be even more relevant when several runs of production are needed or when large amounts of protein are needed. A quick look at the cDNA sequence allows to identify if the sequence contains a high content of rare codons that the bacteria will have a hard time to translate from mRNA into protein.
This step consists in integrating the cDNA into a bacterial expression vector containing a purification tag and inducing the expression of the fusion protein of interest and allowing the protein purification on affinity column via the tag. The tag can be removed if necessary for functional studies.
NB: We perform several runs of tests in order to identify optimal conditions:
In order to identify the optimal conditions for the synthesis of proteins, we need to modify a large number of parameters and we are involved in all stages including codons optimization, vector choice, tag, culture medium, additives, bacterial strains, induction temperatures, IPTG concentrations, purification technique types, renaturation / refolding (dialysis buffer change)… As a leading manufacturer protein, we countinuously strive to improve our processes and it is one of the main strengths of our organization.
NB: the production step is carried out according to customer specifications based on results obtained at the end of step II. Many kinds of purification types are available including tag affinity purification (His, GST, MBP, Sumo), size exclusion chromatography, ion exchange, etc.
Turnaround time :
ProteoGenix has extensive experience in the custom production of proteins in Escherichia coli. We are able to produce any type of protein. Our experience in this industry gives us a large number of solutions and technical advantages that allows us almost always to obtain the required material to:
We also accept protein synthesis orders for very large scale production in fermentors.
E. coli protein production enables to reach high expression yields, at low cost and within a very short period of time because the growth rate of bacteria is much faster than any other system. However, this system is not appropriate when post-translational modifications (PTM) are necessary for the required application.