An expression system and process for the production of Diphtheria toxin polypeptides or mutated forms thereof, such as the toxoid CRM197 polypeptide, in genetically-modified E. coli with high yield is described. The system and process is based on the uncoupling of biomass growth from recombinant protein induction, i.e. using an inducer of protein production that cannot be used as a carbon source for growth by the bacteria. The use of specific components and conditions that improve protein yields are also described.

The present invention relates to a method for producing CRM197 recombinant protein in cells. The method comprises culturing a cell comprising an expression plasmid with a polynucleotide and inducing expression of the CRM197 protein.

The present invention resides in the discovery that the specific interaction between Myeloid Differentiation factor 2 (MD2) and integrin, especially integrin αvβ3, is involved in cellular signaling mediated by MD2-integrin, such as inflammatory response including sepsis. Thus, this invention provides for a novel method for inhibiting integrin signaling by using an inhibitor of MD2-integrin binding, such as a dominant negative mutant of MD2 without integrin-binding capability. A method for identifying inhibitors of MD2-integrin binding is also described. Further disclosed are polypeptides, nucleic acids, host cells, and corresponding compositions for inhibiting MD2-integrin signaling.

Provided herein are lasso peptides libraries, and particularly phage display libraries of lasso peptides. Also provided herein are related methods and systems for producing the libraries and for screening the libraries to identify candidate lasso peptides having desirable properties.

The present invention is a DNA expression vector comprising: a toxP: a mutant toxO that blocks Fe-mediated regulation of gene expression; and a DNA sequence encoding a protein, wherein the toxP and the mutant toxO regulate expression of the DNA segment encoding the protein. It is preferred that DNA expression vectors of the present invention include DNA sequences encoding a signal peptide so that a protein expressed is attached to the signal peptide prior to processing. Novel proteins are produced off of the DNA expression vector of the present invention.

The present invention resides in a method for the manufacture of a disulphide-requiring biopharmaceutical having an element of at least tertiary structure using wild type E. coli.

The present invention is a DNA expression vector comprising: a toxP; a mutant toxO that blocks Fe-mediated regulation of gene expression; and a DNA sequence encoding a protein, wherein the toxP and the mutant toxO regulate expression of the DNA segment encoding the protein. It is preferred that DNA expression vectors of the present invention include DNA sequences encoding a signal peptide so that a protein expressed is attached to the signal peptide prior to processing. Novel proteins are produced off of the DNA expression vector of the present invention.

The present invention is a DNA expression vector comprising: a toxP: a mutant toxO that blocks Fe-mediated regulation of gene expression; and a DNA sequence encoding a protein, wherein the toxP and the mutant toxO regulate expression of the DNA segment encoding the protein. It is preferred that DNA expression vectors of the present invention include DNA sequences encoding a signal peptide so that a protein expressed is attached to the signal peptide prior to processing. Novel proteins are produced off of the DNA expression vector of the present invention.

An expression system and process for the production of Diphtheria toxin polypeptides or mutated forms thereof, such as the toxoid CRM197 polypeptide, in genetically-modified E. coli with high yield is described. The system and process is based on the uncoupling of biomass growth from recombinant protein induction, i.e. using an inducer of protein production that cannot be used as a carbon source for growth by the bacteria. The use of specific components and conditions that improve protein yields are also described.

Provided herein, in some aspects, are methods and compositions for cell-free production of ribonucleic acid.