US 5,223,407 A · Excretion of heterologous proteins from E. Coli

Abstract

A recombinant DNA construct, useful for obtaining excretion from E. coli of a heterologous protein, comprises a coding region in which DNA coding for the heterologous protein in coupled in reading frame with DNA coding for the OmpA signal peptide, to enable secretion of the protein into the periplasm; and a control region linked operably with the coding region to enable expression thereof in an E. coli host, wherein the control region comprises the tac promoter, the lac operator and a consensus ribosome binding site. With this construct, the excretion of heterologous proteins from E. coli can be achieved, consistently and for a wide variety of such proteins, without assistance from carrier proteins or membrane-lytic agent and by using "healthy" hosts rather than mutant strains having defective outer membranes.

Claims19

Dependent claims

An E. coli excretion vector for producing E. coli transformants that excrete heterologous protein from the periplasmic space, said vector comprising a recombinant construct, said construct comprising a coding region in which DNA coding for a heterologous protein sequence is coupled in reading frame with DNA coding for the OmpA signal peptide to enable secretion of protein to the periplasm and whereby said coding region encodes a fusion polypeptide comprising said heterologous protein sequence;

and a control region linked operably with the coding region to enable expression of said coding region in an E. coli host and further enabling excretion of a polypeptide comprising said heterologous protein sequence from the periplasmic space of said host, said control region comprising the tac promoter, the lac operator and a consensus ribosome binding site having the sequence 5'AGGAGGAAAAAATT3'.

Dependencies are inferred from the claim text.

Excretion of heterologous proteins from E. Coli

Life of this patent

Excretion of heterologous proteins from E. Coli

Life of this patent

Abstract

Claims19