The prokaryotic expression system (E. coli expression system) is the earliest developed and most widely used classic expression system in gene expression technology. In recent decades, the E. coli expression system has also been continuously developed and improved, and has been widely used by scientific research and industrial users for various Recombinant protein expression. Compared with other expression systems, it has the characteristics of high expression level of target gene, short culture period, strong anti-pollution ability and relatively low cost.
Medicilon researchers have established a mature E. coli expression and purification service platform to provide expression and purification services including various recombinant proteins and their complexes in E. coli.
Prokaryotic expression system (E. coli expression system) service items:
Recombinant plasmid construction;
Recombinant expression test;
Soluble expression condition test;
Small-scale protein expression and purification (2L medium culture product);
Large-scale protein expression and purification (above 10L).
|Step||Experiment content||Experiment time||Remarks|
|1||Recombinant plasmid construction||for 2 weeks||You can choose different vectors, different tags His, Trx, GST, SUMO…|
|2||Recombinant expression test||1 week||Different expression strains are optional|
|3||Soluble expression test||1-2 weeks||Different inducers, induction temperature, can also be reconsidered based on protein Construction of mutants with structural point mutations or deletion mutations|
|4||Small-scale protein expression and purification||1-2 weeks||In case of inclusion body expression, the time and method will be adjusted|
|5||Large-scale protein expression and purification||According to the expression volume||It can be selected from shake flask or fermentor culture|
You can choose any of the above experimental content, or a combination of related experimental content according to the experimental needs;
During purification, affinity purification, ion column purification, hydrophobic packing purification and molecular sieve purification are used; the characteristics of the protein itself and the choice of expression system will affect the purity and activity of the final protein;