Rna replicon for improving gene expression and use thereof
US-2024417751-A1 · Dec 19, 2024 · US
E. coli deficient in Lon and SulA for gene targeting
US12297239B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-12297239-B2 |
| Application number | US-202318325199-A |
| Country | US |
| Kind code | B2 |
| Filing date | May 30, 2023 |
| Priority date | Aug 17, 2017 |
| Publication date | May 13, 2025 |
| Grant date | May 13, 2025 |
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Abstract
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An expression system and process for the production of Diphtheria toxin polypeptides or mutated forms thereof, such as the toxoid CRM197 polypeptide, in genetically-modified E. coli with high yield is described. The system and process is based on the uncoupling of biomass growth from recombinant protein induction, i.e. using an inducer of protein production that cannot be used as a carbon source for growth by the bacteria. The use of specific components and conditions that improve protein yields are also described.
First claim
Opening claim text (preview).
What is claimed is: 1. An Escherichia coli ( E. coli ) cell comprising a defective sulA gene and a defective lon gene. 2. The E. coli cell of claim 1 , wherein the E. coli cell is a BL21 strain cell. 3. The E. coli cell of claim 2 , wherein the E. coli cell is a BL21 (DE3) strain cell. 4. The E. coli cell of claim 1 , wherein the cell is defective in rhamnose catabolic pathway. 5. The E. coli cell of claim 4 , wherein the cell comprises a defective rhaB gene. 6. The E. coli cell of claim 1 , wherein the defective sulA gene comprises the nucleotide sequence set forth in SEQ ID NO: 10. 7. An expression system comprising the E. coli cell of claim 1 , wherein the E. coli cell comprises a heterologous nucleic acid construct comprising an inducible promoter sequence operably linked to a nucleic acid molecule encoding a polypeptide. 8. The expression system of claim 7 , wherein the inducible promoter is a rhamnose inducible promoter. 9. A method of gene targeting by homologous recombination, the method comprising introducing an in vivo double-strand break into a DNA molecule within an E. coli cell of claim 1 , in the presence of a donor substrate DNA molecule.
Assignees
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Classifications
from fungi, e.g. yeasts · CPC title
Polyols, e.g. glycerin, inositol · CPC title
Sugars · CPC title
Iron; Fe chelators; Transferrin · CPC title
using homologous recombination · CPC title
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- What does patent US12297239B2 cover?
- An expression system and process for the production of Diphtheria toxin polypeptides or mutated forms thereof, such as the toxoid CRM197 polypeptide, in genetically-modified E. coli with high yield is described. The system and process is based on the uncoupling of biomass growth from recombinant protein induction, i.e. using an inducer of protein production that cannot be used as a carbon sou…
- Who is the assignee on this patent?
- Nat Res Council Canada
- What technology area does this patent fall under?
- Primary CPC classification C12N15/70. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue May 13 2025 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 8 related publications on this page (citations in our corpus or others sharing the same primary CPC).