Abstract: The present invention provides expression vectors for use in an inducible coexpression system, capable of controlled induction of expression of each gene product.

Abstract: The present invention provides expression vectors for use in an inducible coexpression system, capable of controlled induction of expression of each gene product.

Abstract: A cell for producing insulin includes an E. coli host cell and a polynucleotide designed for production of insulin disposed within the E. coli host cell. The polynucleotide includes a first inducible promoter and a first coding sequence to be transcribed from the first inducible promoter, the first coding sequence encoding an insulin polypeptide. The polynucleotide also includes a second inducible promoter and a second coding sequence to be transcribed from the second inducible promoter, the second coding sequence encoding one or more of cDsbA, cDsbC, a protein disulfide isomerase, Ervlp, and a chaperone. The first inducible promoter and the second inducible promoter are not responsive to the same inducer. A method of producing insulin includes growing a sample of the cells in a fermentation volume between 0.1 L and 1,000,000 L, adding an inducer of the first inducible promoter to induce expression of the insulin polypeptide, and purifying the insulin polypeptide.

Abstract: The present invention provides host cells having an oxidizing cytoplasm, which are capable of growing to high cell densities and producing recombinant proteins in soluble form. Also provided are methods for growing such host cells to high densities, methods for optionally modulating the growth rates of the host cells, and methods for inducing the host cells to produce the desired gene product in soluble form. Further aspects of the invention relate to host cell preparation and storage methods, which utilize the advantages of producing gene products in host cell cytoplasm to provide methods for long-term storage and/or transport of expressed gene products that are retained in a stable and soluble form within the host cells.

Abstract: The present invention provides expression vectors for use in an inducible coexpression system, capable of controlled induction of expression of each gene product.

Abstract: The present invention provides expression vectors for use in an inducible coexpression system, capable of controlled induction of expression of each gene product.