EP0108045A1
Expired / Lapsed
EP
⏱️ 0.0 years remaining
RecA promoter dependent polypeptide production
Assignee: Monsanto Co
•
Filing: 1983-10-20
•
Granted: —
•
Expires: 2003-10-20
Abstract
Abstract This invention relates to the expression and accumulation in bacterial cells of heterologous polypeptides under the control of the recA promoter/operator region. One such polypeptide comprises a recA/somatostatin fusion polypeptide. This fusion polypeptide has been expressed and accumulated by E. coli with much greater effectiveness than other somatostatin-containing fusion polypeptides. In addition, use of the recA promoter/operator region provides greater induction flexibility than other inducible promoter systems.
Inventors
- 👤 Michael Louis Bittner
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-05 01:57
CPC Classifications
C; C12; C12N; C12N15/00
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
56 words
A method of producing a heterologous polypeptide in bacteria comprising culturing, under suitable conditions that favor the expression of the heterologous polypeptide, suitable bacterial host cells which contain one or more copies of a gene that comprises a recA promoter/operator region and a structural sequence that can be translated by the bacteria into the heterologous polypeptide.
Claim 2
Independent Claim
56 words
A method of producing a heterologous polypeptide in bacteria comprising culturing, under suitable conditions that favor the expression of the heterologous polypeptide, suitable bacterial host cells which contain one or more copies of a gene that comprises a recA promoter/operator region and a structural sequence that can be translated by the bacteria into the heterologous polypeptide.
Claim 3
Dependent Claim
44 words
A method of Claim 1 wherein the suitable conditions that favor the expression of the heterologous polypeptide comprise exposure of the host cells to one or more of the following substances or conditions: nalidixic acid, mitomycin C, ultraviolet radiation, elevated temperature, and thymine starvation.
Claim 4
Dependent Claim
18 words
A method of claim 1 in which the herterologous. polypeptide is produced as part of a fusion polypeptide.
Claim 5
Dependent Claim
21 words
A method of Claim 3 in which the fusion polypeptide comprises the recA polypeptide of E. coli, or a portion thereof.
Claim 6
Dependent Claim
24 words
A method of Claim 3 which further comprises cleaving the heterologous polypeptide from the fusion peptide at a selected amino acid residue or sequence.
Claim 7
Dependent Claim
17 words
A method of Claim 3 in which the heterologous polypeptide comprises the amino acid sequence of somatostatin.
Claim 8
Dependent Claim
28 words
A method of Claim 3 in which the heterologous polypeptide comprises an amino acid sequence or derivative thereof which is a biologically functional analog or fragment of somatostatin.
Claim 9
Dependent Claim
21 words
A method of Claim 4 in which the portion of the recA polypeptide contains more than about 150 amino acid residues.
Claim 10
Dependent Claim
56 words
A method of Claim 8 in which the recA polypeptide fragment is followed by: a. a selected amino acid residue or sequence which provides a site suitable for cleaving the polypeptide at a desired location; and b. a polypeptide chain comprising the amino acid sequence of somatostatin or a biologically functional analog or fragment of somatotatin.
Claim 11
Dependent Claim
65 words
A double-stranded DNA cloning vector comprising, in phase from a first 5' end to a second 3' end of the sense strand thereof: a. a recA promoter/operator region; b. a 5' non-translated region including a ribosome binding site suitable for enhancing translation of a structural gene in host cells; c. a structural sequence capable of translation into a selected heterologous polypeptide in the host cells.
Claim 12
Dependent Claim
65 words
A double-stranded DNA cloning vector comprising, in phase from a first 5' end to a second 3' end of the sense strand thereof: a. a recA promoter/operator region; b. a 5' non-translated region including a ribosome binding site suitable for enhancing translation of a structural gene in host cells; c. a structural sequence capable of translation into a selected heterologous polypeptide in the host cells.
Claim 13
Dependent Claim
37 words
The cloning vector of Claim 10 in which the DNA sequence which codes for the heterologous polypeptide is fused to a DNA sequence which codes for a polypeptide which is endogenous with respect to the host cell.
Claim 14
Dependent Claim
19 words
The cloning vector of Claim 11 in which the endogenous polypeptide is the recA polypeptide or a portion thereof.
Claim 15
Dependent Claim
21 words
The cloning vector of Claim 11 in which the heterologous polypeptide is somatostatin or a biologically functional analog or fragment thereof.
Claim 16
Dependent Claim
31 words
A double-stranded DNA cloning vector comprising: a. a recA promoter operator region; and b. a 5' non-translated region which is derived from one or more genes other than the recA gene.
Claim 17
Dependent Claim
31 words
A double-stranded DNA cloning vector comprising: a. a recA promoter operator region; and b. a 5' non-translated region which is derived from one or more genes other than the recA gene.
Claim 18
Dependent Claim
15 words
Plasmid pMON2506 characterized as shown by the restriction map in FIG. 4 of the drawings.
Claim 19
Dependent Claim
15 words
Plasmid pMON2506 characterized as shown by the restriction map in FIG. 4 of the drawings.
Claim 20
Dependent Claim
15 words
Plasmid pMON2507 characterized as shown by the restriction map in FIG. 5 of the drawings.
Claim 21
Dependent Claim
15 words
Plasmid pMON2507 characterized as shown by the restriction map in FIG. 5 of the drawings.
Claim 22
Dependent Claim
18 words
A culture of cells having the relevant characteristics of a culture of cells having ATCC accession number 39211.
Claim 23
Dependent Claim
18 words
A culture of cells having the relevant characteristics of a culture of cells having ATCC accession number 39211.
Claim 24
Dependent Claim
18 words
A culture of cells having the relevant characteristics of a culture of cells having ATCC accession number 39212.
Claim 25
Dependent Claim
18 words
A culture of cells having the relevant characteristics of a culture of cells having ATCC accession number 39212.
Claim 26
Dependent Claim
17 words
A culture of cells replicated or derived from a culture of cells having ATCC accession number 39211.
Claim 27
Dependent Claim
17 words
A culture of cells replicated or derived from a culture of cells having ATCC accession number 39211.
Claim 28
Dependent Claim
17 words
A culture of cells replicated or derived from a culture of cells having ATCC accession number 39212.
Claim 29
Dependent Claim
17 words
A culture of cells replicated or derived from a culture of cells having ATCC accession number 39212.
Claim 30
Dependent Claim
32 words
A polypeptide comprising : a. the amino acid sequence of recA, or a portion or derivative thereof; b. the amino acid sequence of somatostatin, or a biologically functional analog or fragment thereof.
Claim 31
Dependent Claim
32 words
A polypeptide comprising : a. the amino acid sequence of recA, or a portion or derivative thereof; b. the amino acid sequence of somatostatin, or a biologically functional analog or fragment thereof.
Claim 32
Dependent Claim
20 words
A polypeptide of Claim 21, containing more than about 150 amino acid residues of the amino acid sequence of recA.
Claim 33
Dependent Claim
25 words
A polypeptide of Claim 22, containing a selected amino acid residue or sequence which provides a method of cleaving the polypeptide at a desired location.
References Cited (Backward Citations)
GB2007676A
Cited
EP0036776A2
Cited
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.