Expression system for product manufacturing
AssigneeNovel Biotechnology USA, Inc.
InventorsSanthosh Thomas Kallivalappil, Matthew Weinstock
Life of this patent
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Abstract
The present disclosure provides engineered bacterial cells having one or more genetic modifications that result in increased production of expression products, such as increased production of plasmids with reduced occurrence of plasmid concatemers or multimers. Also disclosed herein are expression systems for product manufacturing in media having reduced antibiotic concentration, as well as methods of making cells for use in the expression systems. The cells and expression systems increase the growth rate and production yields of biomolecules produced within the host bacterial host cells without the use of antibiotics, and can produce the product with little to no endotoxins present.
Claims18
A genetically-modified Vibrio natriegens bacterial cell having a genetic modification with respect to a parent Vibrio natriegens bacterial cell of a strain selected from the group consisting of:
- Vibrio natriegens bacterial strain deposited under accession number NCIMB 857, Vibrio natriegens bacteria strain deposited under accession number ATCC 14048, Vibrio natriegens bacteria strain deposited under accession number DSM 759, and Vibrio natriegens bacteria strain deposited under accession number NBRC 15636;
- wherein the genetic modification is a modification of a chromosome 1 gene that has a polynucleotide sequence of SEQ ID NO:
- 1 prior to the modification and wherein the modification results in reduced expression of a polypeptide encoded by the polynucleotide sequence of SEQ ID NO:
- 1, as compared to a level of expression of the polypeptide encoded by the polynucleotide sequence of SEQ ID NO:
- 1 in the parent Vibrio natriegens bacterial cell.
A method of making a genetically-modified Vibrio natriegens bacterial cell, the method comprising:
- (a) obtaining a parent Vibrio natriegens bacterial cell of a strain selected from the group consisting of:
- a Vibrio natriegens bacterial strain deposited under accession number NCIVB 857, a Vibrio natriegens bacteria strain deposited under accession number ATCC 14048, a Vibrio natriegens bacteria strain deposited under accession number DSM 759, and a Vibrio natriegens bacteria strain deposited under accession number NBRC 15636;
- and
- (b) performing a modification of a chromosome 1 gene of the parent Vibrio natriegens bacterial cell that has a polynucleotide sequence of SEQ ID NO:
- 1 prior to the modification, thereby generating the genetically-modified Vibrio natriegens bacterial cell.
A method of replicating a template plasmid with reduced occurrence of plasmid multimers or concatemers in a genetically-modified Vibrio bacterial cell, the method comprising:
- (a) obtaining the genetically-modified Vibrio bacterial cell, wherein the genetically-modified Vibrio bacterial cell comprises a genetic modification with respect to a parent Vibrio natriegens bacterial cell of a strain selected from the group consisting of:
- a Vibrio natriegens bacterial strain deposited under accession number NCIVIB 857, a Vibrio natriegens bacteria strain deposited under accession number ATCC 14048, a Vibrio natriegens bacteria strain deposited under accession number DSM 759, and a Vibrio natriegens bacteria strain deposited under accession number NBRC 15636;
- wherein the genetic modification is a modification of a chromosome 1 gene that has a polynucleotide sequence of SEQ ID NO:
- 1 prior to the modification and wherein the modification results in reduced expression of a polypeptide encoded by the polynucleotide sequence of SEQ ID NO:
- 1, as compared to a level of expression of the polypeptide encoded by the polynucleotide sequence of SEQ ID NO:
- 1 in the parent Vibrio natriegens bacterial cell;
- and
- (b) transforming the template plasmid into the genetically-modified Vibrio bacterial cell, thereby replicating the template plasmid in the genetically-modified Vibrio bacterial cell;
- wherein the template plasmid is replicated in the genetically-modified Vibrio bacterial cell with reduced levels of plasmid multimers or concatemers, relative to an amount of plasmid multimers or concatemers produced in the same amount of time upon transformation of the template plasmid into a bacterial cell of a parent Vibrio natriegens bacterial cell lacking the genetic modification.
Dependencies are inferred from the claim text.