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US 9,617,335 B2Granted
Inducible coexpression system
AssigneeAbSci LLC
InventorsSean McClain, Mark Valasek
Life of this patent
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Abstract
The present invention is an inducible coexpression system, capable of controlled induction of expression of each gene product.
Claims30
An Escherichia coli host cell comprising two or more types of expression constructs,
- wherein the expression construct of each type comprises an inducible promoter and a polynucleotide sequence encoding a gene product to be transcribed from the inducible promoter;
- and
- at least one of said inducible promoters is responsive to an inducer that is different than the inducer of another of said inducible promoters;
- and
- each inducible promoter is selected from the group consisting of:
- an L-arabinose-inducible promoter, a propionate-inducible promoter, a rhamnose-inducible promoter, and a D-xylose-inducible promoter;
- and
- wherein the host cell has a reduced level of gene function of at least one gene encoding a protein that metabolizes an inducer of at least one said inducible promoter;
- and
- wherein the host cell has a reduced level of gene function of a gene that encodes a reductase;
- and
- at least one of said gene products forms a multimer with another of said gene products;
- and
- at least one of said gene products is a polypeptide that lacks a signal peptide.
11 more elements
An Escherichia coli host cell comprising two or more types of expression constructs,
- wherein the expression construct of each type comprises an inducible promoter and a polynucleotide sequence encoding a gene product to be transcribed from the inducible promoter;
- and
- at least one of said inducible promoters is responsive to an inducer that is different than the inducer of another of said inducible promoters;
- and
- each inducible promoter is selected from the group consisting of:
- an L-arabinose-inducible promoter, a propionate-inducible promoter, a rhamnose-inducible promoter, and a D-xylose-inducible promoter;
- and
- wherein the host cell has a reduced level of gene function of at least one gene encoding a protein that metabolizes an inducer of at least one said inducible promoter;
- and
- wherein the host cell has a reduced level of gene function of a gene that encodes a reductase;
- and
- at least one of said gene products is a polypeptide that lacks a signal peptide.
9 more elements
An Escherichia coli host cell comprising two or more types of expression constructs,
- wherein the expression construct of each type comprises an inducible promoter and a polynucleotide sequence encoding a gene product to be transcribed from the inducible promoter;
- and
- at least one of said inducible promoters is responsive to an inducer that is different than the inducer of another of said inducible promoters;
- and
- each inducible promoter is selected from the group consisting of:
- an L-arabinose-inducible promoter, a propionate-inducible promoter, a rhamnose-inducible promoter, and a D-xylose-inducible promoter;
- and
- wherein the host cell has a reduced level of gene function of at least one gene encoding a protein that metabolizes an inducer of at least one said inducible promoter;
- and
- at least one of said gene products forms a multimer with another of said gene products;
- and
- at least one gene product is selected from the group consisting of:
- (a) an immunoglobulin heavy chain;
- (b) an immunoglobulin light chain;
- and (c) a fragment of any of (a)-(b).
12 more elements
Dependencies are inferred from the claim text.