Methods of cloning nucleic acids or producing proteins in a low endotoxin organism
AssigneeTelesis Bio Inc.
InventorsMatthew T Weinstock, Daniel G. Gibson, Daniel Strimling
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Abstract
The invention provides engineered Vibrio sp. organisms that comprise a genetic modification to either or both of the lpxL and/or lpxM genes. The organisms score substantially lower in an in vitro endotoxin assay versus the unmodified or wild type organism. The organisms preserve substantially the growth rate of the corresponding unmodified organisms. The organisms can also have an exogenous nucleic acid cloned in the organism, or an exogenous nucleic acid encoding a protein, polypeptide, or peptide expressed by the organism, and optionally secreted from the organism.
Claims18
A method of cloning a nucleic acid or of producing a protein comprising:
- culturing a recombinant Vibrio sp. organism comprising:
- an exogenous nucleic acid sequence for cloning by the organism, or an exogenous nucleic acid sequence encoding a heterologous protein or peptide for production by the organism;
- a genetic modification selected from deletion, inactivation, or disruption of the lpxL gene or the lpxM gene, wherein the organism produces substantially less endotoxin compared to a corresponding organism not comprising the deletion, inactivation, or disruption and cultivated under the same conditions;
- and
- wherein the recombinant Vibrio sp. organism exhibits a growth rate of at least 60% of the growth rate of the corresponding organism when cultivated under the same conditions;
- wherein the organism has an endotoxin level of less than 50 EU/ml of purified lipopolysaccharide;
- and
- harvesting the nucleic acid sequence cloned by the organism, or the protein or peptide produced by the organism.
Dependencies are inferred from the claim text.