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US 12,404,524 B2Granted
Expression vector
AssigneeTerra Bioworks, Inc.
InventorsRobert Joseph Stankey, David Mead
Life of this patent
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Abstract
Disclosed herein are recombinant methods of activating expression of one or more biosynthetic gene clusters comprising more than one gene, the method comprising a recombinant DNA expression vector that possess two opposable inducible promoters that drives expression of a biosynthetic gene cluster exogenously from outside of the cluster to produce polyketides or non-ribosomal peptides in a heterologous host.
Claims16
A method of expressing a polypeptide from an insert nucleic acid comprising a biosynthetic gene cluster, the method comprising:
- providing a nucleic acid comprising a biosynthetic gene cluster;
- providing an expression vector comprising a first promoter and a second promoter flanking a cloning site, wherein the first promoter and the second promoter direct transcription toward each other and in opposite directions;
- cloning an insert nucleic acid comprising the biosynthetic gene cluster at said cloning site to provide a recombinant expression vector comprising said insert nucleic acid;
- transforming a host cell with the recombinant expression vector;
- expressing a polypeptide from the biosynthetic gene cluster;
- and
- detecting the expression of the polypeptide using a functional selection or screen that is specific for the polypeptide or for a product that is indicative of the polypeptide.
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Dependencies are inferred from the claim text.