Method for producing a recombinant protein
AssigneeThe University of Tennessee Research Corporation
InventorsRonald K. Taylor, Joel A. Peek
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Abstract
A method to inhibit the function of a class of bacterial periplasmic oxidoreductase enzymes, exemplified by TcpG of Vibrio cholerae, DsbA of E. coli, and Por of Haemophilus influenzae, is disclosed. A screening method to determine whether a chemical inhibits the function of the oxidoreductase enzymes is disclosed. Mutant bacteria lacking the ability to produce a functional periplasmic oxidoreductase and a cloned expression vector overproducing TcpG are also disclosed.
Claims18
A method for producing a recombinant protein comprising transforming a host bacterium with a first gene encoding a recombinant protein, transforming said host bacterium with a second gene encoding a bacterial periplasmic protein selected from the group consisting of Por, TcpG, and DsbA, and culturing said host bacterium to produce said recombinant protein, which periplasmic protein catalyzes the formation of disulfide bonds between cysteine residues, wherein said host bacterium is selected from the group consisting of E. coli, Haemophilus influenzae, Salmonella typhimurium, Bacillus brevis, Legionella, Erwinia chryantum, Streptococcus, and Staphylococcus.
A method for producing a recombinant protein comprising inserting a first nucleic acid sequence under the control of a promoter into the genome of a host bacterium, inserting into said host genome a second nucleic acid sequence under the control of a promoter, which second nucleic acid sequence encodes a bacterial periplasmic enzyme which catalyzes the formation of disulphide bonds between two cysteine residues, which bacterial periplasmic enzyme is selected from the group consisting of Por, TcpG, and DsbA, and co-expressing the first and second nucleic acid sequences, wherein said host bacterium is selected from the group consisting of E. coli, Haemophilus influenzae, Salmonella typhimurium, Bacillus brevis, Legionella, Erwinia chrysantum, Streptococcus, and Staphylococcus.
Dependencies are inferred from the claim text.