E. coli Expression Services
Our E. coli Expression System offers flexibility you need to express and produce the recombinant protein of your choice.
BPS will clone your insert into an expression vector. Either a standard subcloning or a PCR cloning will be used to transfer the gene of interest into the expression vector. Clones will be screened for the correct restriction fragments and sequenced.
The expression vector containing the gene of interest will be transformed into an expression strain of E. coli for protein production. Three to five individual clones will be screened in a small-scale induction experiment and analyzed by SDS/PAGE for production of a protein of the expected size. If desired, a Western blot can be carried out to verify immunoreactivity with your antisera, or with an anti-affinity tag antisera.
Optimization of Expression
The expression of the recombinant protein can be optimized by doing a time course of induction, altering induction temperature or screening growth media. For each test, the recombinant protein will be analyzed by SDS-PAGE and/or Western blot to determine optimal conditions for protein production.
A small-scale culture (10-100 ml) from one isolate of the strain producing the recombinant protein will be induced and the expressed protein will be characterized for cellular location, solubility, and accessibility of purification tags. In addition, the yield per unit volume will be estimated. Using the results of these experiments, a strategy for producing and purifying the protein will be made.
Using shake flasks the strain expressing the recombinant protein can be scaled up between one and several liter volumes. You will receive the cell pellets and a complete project report with the SDS-PAGE and/or Western blot analysis that document protein production.
BPS will purify your recombinant protein(s). We offer a variety of services from inclusion body preparation to affinity column purification.