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US 2011/0033893 A1Published application
Improved methods for protein production
OwnerRobert L. Garcea, Anton Middelberg +1inventors
InventorsRobert L. Garcea, Anton Middelberg, Yap Pang Chuan
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Abstract
Methods for low cell-density bacterial protein expression that can achieve levels of up to 180 mg/l using a simple and low cost strategy. Full codon optimization is unnecessary to improve expression of viral genes rich in E. coli rare codons. Using a strategically modified host cell provides a simpler and cheaper alternative.
Claims28
A method for producing a protein comprising:
- a) culturing bacteria until the bacteria reach stationary phase, wherein the bacteria express a recombinant nucleic acid molecule encoding the protein and wherein expression of the protein is inducible;
- b) cooling the culture to about 25° C.;
- c) inducing protein expression by adding an induction agent;
- and
- d) culturing the bacteria at about 25° C.
2 more elements
A method for producing a GST-HPV L1 fusion protein or a MPV VP1 protein comprising:
- a) culturing E. coli until the E. coli reach stationary phase, wherein the E. coli express a recombinant nucleic acid molecule encoding the GST-HPV L1 fusion protein or the MPV VP1 protein and wherein expression of the protein is inducible by IPTG;
- b) cooling the culture to about 25° C.;
- c) inducing protein expression by adding an IPTG;
- and
- d) culturing the bacteria at about 25° C.
2 more elements
Dependencies are inferred from the claim text.