E.coli expression takes 6 to 8 weeks from subcloning to expression in 2 phases starting at $2,200.
Subcloning into Expression Vector
- Amplification/isolation of the gene of interest (GOI) from a client-supplied construct and subcloning into a suitable transfer vector with His-Tag (or GST-tag).
- Sequence confirmation of insert.
- Mini-induction to over-express the target protein.
- Test for recombinant protein by SDS-PAGE and Western Blot (customer must provide appropriate antibodies to protein of interest(POI), if desired).
Estimated time: 3-5 weeks
Expansion and Purification
- One liter of bacterial culture will be induced with IPTG and harvested for protein purification.
- Ni-NTA purification of protein with 6X His-tags or glutathione for GST-tagged proteins using beads either under native or denatured conditions.
Estimated time: 2 weeks
- 5 μg of transfer vector
- Recombinant Protein
- Yield ranges from 0.5 – 50 mg/L of culture with average yield of 3 mg/L. Solubility is not guaranteed.