EP0036259A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Method,vectors and organisms for transported cloned heterologous gene products in bacillus subtilis and E.coli
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1981-03-02
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Granted: —
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Expires: 2001-03-02
Abstract
Abstract A method and a cloning vector are described for the controlled accumulation of cloned heterologous gene products in Bacillus subtilis. The cloning vector is capable of being replicated in B. subtilis and includes the heterologous gene located and oriented such as to be under the control of an operator, promoter, and ribosomal binding site sequence. The gene codes for a protein which is under the control of a transport mechanism by which the protein is secreted by the B. subtilis. The gene product is recovered from the growth medium for the B. subtilis. The cloning vector is also capable of similar use in other bacteria such asE. coli.
Inventors
- 👤 Shing Chang
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-27 08:13
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/75; C12N15/11; C12N15/62; C12N15/625; C12N9/00; C12N9/14; C12N9/78; C12N9/86; C07; C07K; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/036; C07K2319/61; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 51
Claim 1
Independent Claim
98 words
A method for producing a predetermined protein comprising providing growth conditions in a growth medium for gram positive bacteria containing cloning vectors each having a gene therein for said predetermined protein, said predetermined protein being non-indigenous to the gram positive bacteria, said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal binding site sequence, said predetermined protein being under the control of a transport mechanism by which said predetermined protein is secreted by said gram positive bacteria, and recovering said predetermined protein from said growth medium.
Claim 2
Independent Claim
98 words
A method for producing a predetermined protein comprising providing growth conditions in a growth medium for gram positive bacteria containing cloning vectors each having a gene therein for said predetermined protein, said predetermined protein being non-indigenous to the gram positive bacteria, said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal binding site sequence, said predetermined protein being under the control of a transport mechanism by which said predetermined protein is secreted by said gram positive bacteria, and recovering said predetermined protein from said growth medium.
Claim 3
Dependent Claim
12 words
A method according to Claim 1 wherein said bacteria are Bacillus subtilis.
Claim 4
Dependent Claim
27 words
A method according to Claim 1 wherein said gene for said predetermined protein is under the control of the beta-lactamase operator, promoter, and ribosomal binding site sequence.
Claim 5
Dependent Claim
12 words
A method according to Claim 1 wherein said predetermined protein is beta-lactamase.
Claim 6
Dependent Claim
20 words
A method according to Claim 1 wherein said transport function is provided for by a portion of the beta-lactamase gene.
Claim 7
Dependent Claim
20 words
A method according to Claim 1 wherein said transport function is provided for by a portion of the alpha-amylase gene.
Claim 8
Dependent Claim
14 words
A method according to Claim 1 wherein said predetermined protein is a mammalian protein.
Claim 9
Dependent Claim
14 words
A method according to Claim 1 wherein said predetermined protein is a mammalian hormone.
Claim 10
Independent Claim
73 words
A method for producing a predetermined protein non-indigenous to Bacillus subtilis comprising providinq growth conditions in a growth medium for Bacillus subtilis bacteria containing cloning vectors each having a gene therein for said predetermined protein, said gene being located and oriented in said vector such as to be under the control of an operator, promoter and ribosomal binding site sequence, and recovering said predetermined protein as a product of said Bacillus subtilis bacteria.
Claim 11
Independent Claim
73 words
A method for producing a predetermined protein non-indigenous to Bacillus subtilis comprising providinq growth conditions in a growth medium for Bacillus subtilis bacteria containing cloning vectors each having a gene therein for said predetermined protein, said gene being located and oriented in said vector such as to be under the control of an operator, promoter and ribosomal binding site sequence, and recovering said predetermined protein as a product of said Bacillus subtilis bacteria.
Claim 12
Dependent Claim
53 words
A method according to Claim 9 wherein said gene for said predetermined protein is additionally located and oriented in said vector such as to be under the control of a transport mechanism by which said predetermined protein is secreted by said Bacillus subtilis bacteria and recoverinq said predetermined. protein from said growth medium.
Claim 13
Independent Claim
29 words
A method according to Claims 9 or 10 wherein said gene for said predetermined protein is under the control of the beta-lactamase operator, promoter, and ribosomal binding site sequence.
Claim 14
Dependent Claim
15 words
. A method according to Claims 9 or 10 wherein said predetermined protein is beta-lactamase.
Claim 15
Dependent Claim
21 words
A method according to Claim 10 wherein said ' transport function is provided for by a portion of the beta-lactamase gene.
Claim 16
Dependent Claim
20 words
A method according to Claim 10 wherein said transport function is provided for by a portion of the. alpha-amylase gene.
Claim 17
Independent Claim
16 words
A method according to Claims 9 or 10 wherein said predetermined protein is a mammalian protein.
Claim 18
Independent Claim
16 words
A method according to Claims 9 or 10 wherein said predetermined protein is a mammalian hormone.
Claim 19
Dependent Claim
80 words
A cloning vector for producing a predetermined protein through expression by B. subtilis, comprising a vector which is capable of autonomous replication in B. subtilis and having a gene therein for said predetermined protein, said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal binding site sequence, said predetermined protein being under the control of a transport mechanism by which said predetermined protein is secreted by B. subtilis.
Claim 20
Dependent Claim
80 words
A cloning vector for producing a predetermined protein through expression by B. subtilis, comprising a vector which is capable of autonomous replication in B. subtilis and having a gene therein for said predetermined protein, said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal binding site sequence, said predetermined protein being under the control of a transport mechanism by which said predetermined protein is secreted by B. subtilis.
Claim 21
Dependent Claim
29 words
A cloning vector according to Claim 17 comprising a plasmid selected from the qroup consisting of: pC221, pUB110, pC194, pUB112, pT127, pOG2165, pOG2110, pCS1006, pOG1196, pCS832, and their derivatives.
Claim 22
Dependent Claim
28 words
A cloning vector according to Claim 17 wherein said gene for said predetermined protein is under the control of the beta-lactamase operator, promoter, and ribosomal binding site sequence.
Claim 23
Dependent Claim
13 words
A cloning vector according to Claim 17 wherein said predetermined protein is beta-lactamase.
Claim 24
Dependent Claim
21 words
A cloning vector according to Claim 17 wherein said transport function is provided for by a portion of the beta-lactamase gene.
Claim 25
Dependent Claim
23 words
A cloning vector according to Claim 17 wherein said transport function is provided for by a portion of the eukaryotic insulin signal peptide.
Claim 26
Dependent Claim
21 words
A cloning vector according to Claim 17 wherein said transport function is provided for by a portion of the alpha-amylase gene.
Claim 27
Dependent Claim
15 words
A cloning vector according to Claim 17 wherein said predetermined protein is a mammalian protein.
Claim 28
Dependent Claim
15 words
A cloning vector according to Claim 17 wherein said predetermined protein is a mammalian hormone.
Claim 29
Dependent Claim
18 words
A cloning vector comprising a plasmid selected from the group consisting of pOG211Q, pOG1196, pOG2165, and their derivatives.
Claim 30
Dependent Claim
18 words
A cloning vector comprising a plasmid selected from the group consisting of pOG211Q, pOG1196, pOG2165, and their derivatives.
Claim 31
Independent Claim
96 words
A method for creating a microorganism capable of producing a predetermined protein through expression, comprising, forming a cloning vector capable of autonomous replication is B. subtilis and having a gene therein for said predetermined protein, said predetermined protein being non-indigenous to B. subtilis, said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal binding site sequence, said predetermined protein being under the control of a transport mechanism hv which said predetermined protein is secreted by B. subtilis, and introducing said vector to B. subtilis.
Claim 32
Independent Claim
96 words
A method for creating a microorganism capable of producing a predetermined protein through expression, comprising, forming a cloning vector capable of autonomous replication is B. subtilis and having a gene therein for said predetermined protein, said predetermined protein being non-indigenous to B. subtilis, said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal binding site sequence, said predetermined protein being under the control of a transport mechanism hv which said predetermined protein is secreted by B. subtilis, and introducing said vector to B. subtilis.
Claim 33
Dependent Claim
27 words
A method according to Claim 27 wherein said gene for said predetermined protein is under the control of the beta-lactamase operator, promoter, and ribosomal binding site sequence.
Claim 34
Dependent Claim
11 words
A method according to Claim 27 wherein said protein is beta-lactamase.
Claim 35
Dependent Claim
20 words
A method according to Claim 27 wherein said transport function is provided for by a portion of a beta-lactamase qene..
Claim 36
Dependent Claim
20 words
A method according to Claim 27 wherein said transport function is provided for by a portion of the alpha-amylase gene.
Claim 37
Dependent Claim
95 words
A microorganism capable of producing a predetermined protein through expression, comprising: a host microorganism, a cloninq vector capable of autonomous replication in said host; said vector having a gene therein for said predetermined protein; said gene being for a predetermined protein which is non-indigenous to said host microorganism; said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal bindinq site sequence ; said predetermined protein being under the control of a transport mechanism by which said protein is secreted by said host microorganism.
Claim 38
Dependent Claim
95 words
A microorganism capable of producing a predetermined protein through expression, comprising: a host microorganism, a cloninq vector capable of autonomous replication in said host; said vector having a gene therein for said predetermined protein; said gene being for a predetermined protein which is non-indigenous to said host microorganism; said gene being located and oriented in said vector such as to be under the control of an operator, promoter, and ribosomal bindinq site sequence ; said predetermined protein being under the control of a transport mechanism by which said protein is secreted by said host microorganism.
Claim 39
Dependent Claim
22 words
A microorganism accordinq to Claim 32 wherein said host microogranism is selected from the group consisting of R- subtilis and E. coli.
Claim 40
Dependent Claim
32 words
A microorganism according to Claim 32 wherein said cloninq vector comprises a plasmid selected from the group consistina of pC221, pUFB110, pC194, pUBl12, pT127, pOG2165, pOG2110, pCS1006, pOG1196, pCS832, and their derivatives.
Claim 41
Dependent Claim
27 words
A microorganism according to Claim 32 wherein said gene or said predetermined protein is under the control of the beta-lactamase operator, promoter, and ribosomal binding site sequence.
Claim 42
Dependent Claim
14 words
A microorganism according to Claim 32 wherein said predetermined protein is a mammalian protein.
Claim 43
Dependent Claim
14 words
A microorganism according to Claim 32 wherein said predetermined protein is a mammalian hormone.
Claim 44
Dependent Claim
12 words
A microoraanism according to Claim 32 wherein said predetermined protein is beta-lactamase.
Claim 45
Dependent Claim
20 words
A microorganism accordinq to Claim 32 wherein said transport function is provided for by a portion of the beta-lactamase gene.
Claim 46
Dependent Claim
22 words
A microorganism according to Claim 32 wherein said transport function is provided for by a portion of the eukarvotic insulin siqnal peptide.
Claim 47
Dependent Claim
22 words
A microorqanism according to Claim 32 wherein said transport function is provided for by a portion of the alpha-amylase q p ne.
Claim 48
Dependent Claim
21 words
A microorganism according to Claim 32 wherein said bifunctional plasmid possesses unique sites for restriction enzymes SStI, HindIII, PstI., and BglII.
Claim 49
Dependent Claim
11 words
A protein produced in accordance w.ith the method of Claim 1.
Claim 50
Dependent Claim
11 words
A protein produced by a microorganism in accordance with Claim 32.
Claim 51
Dependent Claim
26 words
A transformant bacterial culture cloned from one or more bacteria according to Claim 32, the members of said culture being capable of expressing said predetermined protein.
References Cited (Backward Citations)
No citation records indexed.
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