EP0133756A2
Expired / Lapsed
EP
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Vector for expression of polypeptides
Assignee: Genex Corp
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Filing: 1984-07-06
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Granted: —
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Expires: 2004-07-06
Abstract
Abstract A replicable plasmidic expression vector, capable of high levels of expression and secretion of polypeptides in a bacillus is disclosed. The vector contains a DNA sequence comprising the promoter and regulatory regions which control expression and secretion of proteases in a bacillus operably liked to a DNA sequence encoding the amino acid sequence of a polypeptide. The expression vector is particularly useful in the production of B. amyloliquefaciens proteases in B. subtilis. The method facilitates the isolation of proteases from various bacilli.
Inventors
- 👤 Vasantha Nagarajan
- 👤 Carl D. B. Banner
- 👤 Craig Stuart Rhodes
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-27 20:58
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/75; C07; C07K; C07K14/00; C07K14/195; C07K14/305; C07K14/31; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/52; C12N9/54; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/036; C07K2319/61; C07K2319/70; C07K2319/705
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
64 words
A replicable plasmidic expression vector capable of directing expression and secretion of a polypeptide in a bacillus transformed therewith, which comprises: (a) a replicon; (b) a DNA sequence comprising the promotor and regulatory regions which control expression and secretion of a bacillus protease; and (c) a DNA sequence encoding the amino acid sequence of a polypeptide, operably linked to said promotor and regulatory region.
Claim 2
Dependent Claim
64 words
A replicable plasmidic expression vector capable of directing expression and secretion of a polypeptide in a bacillus transformed therewith, which comprises: (a) a replicon; (b) a DNA sequence comprising the promotor and regulatory regions which control expression and secretion of a bacillus protease; and (c) a DNA sequence encoding the amino acid sequence of a polypeptide, operably linked to said promotor and regulatory region.
Claim 3
Dependent Claim
26 words
A replicable plasmidic expression vector a claimed in claim 1 wherein the DNA sequence comprising the said promotor and regulatory regions are derived from B. amyloliquefaciens.
Claim 4
Dependent Claim
40 words
A replicable plasmidic expression vector as claimed in claim 1 or claim 2 wherein the polypeptide which is encoded is the protease whose expression and secretion in its cell of origin is controlled by the said promotor and regulatory regions.
Claim 5
Dependent Claim
44 words
A replicable plasmidic expression vector as claimed in claim 1 or claim 2 wherein the polypeptide which is encoded is a heterologous polypeptide whose expression and secretion in its cell or origin is not controlled by the said protease promotor and regulatory regions selected.
Claim 6
Dependent Claim
30 words
A replicable plasmidic expression vector as claimed in any of claims 1, 2 or 3 wherein the encoding DNA sequence is that coding for a protease derived from B. amyloliquefaciens.
Claim 7
Dependent Claim
35 words
A replicable plasmidic expression vector as claimed in claim 1 or claim 4 wherein the strucutural gene which is fused to the promotor and regulatory region comprises a gene encoding a protein having prorennin activity.
Claim 8
Dependent Claim
36 words
A replicable plasmidic expression vector as claimed in claim 1 or claim 4, wherein the structural gene which is fused to the promotor and regulatory region comprises a gene encoding a protein having protein A activity.
Claim 9
Dependent Claim
49 words
A vector comprising a replicable plasmid containing the promotor and regulatory region of a gene selected from apr[BamP] and npr[BamP] and a sequence of deoxynucleotides, adjacent the 3' end of the signal peptide coding sequence, which comprise an endonuclease restriction site not naturally present in apr[BamP] or p pr[BamP].
Claim 10
Dependent Claim
49 words
A vector comprising a replicable plasmid containing the promotor and regulatory region of a gene selected from apr[BamP] and npr[BamP] and a sequence of deoxynucleotides, adjacent the 3' end of the signal peptide coding sequence, which comprise an endonuclease restriction site not naturally present in apr[BamP] or p pr[BamP].
Claim 11
Dependent Claim
19 words
A plasmid selected from the group consisting of pGx 2109, pGx 2110, pGx 2111, pGx 2134 and pGx 2138.
Claim 12
Dependent Claim
19 words
A plasmid selected from the group consisting of pGx 2109, pGx 2110, pGx 2111, pGx 2134 and pGx 2138.
Claim 13
Dependent Claim
32 words
A bacillus transformed by a vector or plasmid as claimed in any of the preceding claim which is different from the bacillus from which the promotor and regulatory DNA sequence is taken.
Claim 14
Dependent Claim
15 words
B. subtilis transformed by a vector as claimed in any of claims 1 to 10.
Claim 15
Dependent Claim
23 words
A method for producing a microorganism for the expression of a polypeptide which comprises transforming a Bacillus with vector as-claimed in claim 1.
Claim 16
Independent Claim
60 words
A method of producing a polypeptide which comprises: (a) transforming a bacillus with a replicable plasmidic expression vector as claimed in any of claims 1 to 10; (b) growing the transformed bacilli in a culture medium and subjecting them to conditions under which the polypeptide is expressed and secreted into the medium; and (c) recovering the polypeptide from the medium.
References Cited (Backward Citations)
No citation records indexed.
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