EP0063494A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Method for producing protein from a microorganism, microorganisms for use in such method and creation thereof, vectors for use in said creation, and protein produced thereby, and transformant culture derived from said microorganisms
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1982-04-20
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Granted: —
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Expires: 2002-04-20
Abstract
Abstract The invention discloses an improved method and cion- ing vectors useful for the production of cloned heterologous gene products in B. subtilis. Use of the method and vectors allows the host to produce the heterologous gene product as a single unfused peptide having no extraneous amino acids attached that will accumulate within transformed host organisms. The method comprises providing growth conditions in a growth media for B. subtilis bacteria containing plasmids capable of replication in B. subtilis or capable of integrating into the bacterial chromosome of B. subtilis, said plasmids having a heterologous gene therein coding for the predetermined protein and capable of expession in B. subtilis, said heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene, and there being no translational initiation codon sequences prior to said heterologous gene, and optionally recovering the resulting predetermined protein.
Inventors
- 👤 Shing Chang
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-27 16:21
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C07K14/565; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/75; C07K2319/00; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
130 words
A method for producing a predetermined protein that will accumulate within a transformed host comprising providing growth conditions in a growth media for B. subtilis bacteria containing plasmids capable of replication in B. subtilis or capable of integrating into the bacterial chromosome of B. subtilis, said plasmids having a heterologous gene therein coding for the predetermined protein and capable of expression in B. subtilis, said heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene,and there bei-n g- no translational initiation codon sequences prior to said heterologous gene, 'and optionally recovering the resulting predetermined protein.
Claim 2
Independent Claim
130 words
A method for producing a predetermined protein that will accumulate within a transformed host comprising providing growth conditions in a growth media for B. subtilis bacteria containing plasmids capable of replication in B. subtilis or capable of integrating into the bacterial chromosome of B. subtilis, said plasmids having a heterologous gene therein coding for the predetermined protein and capable of expression in B. subtilis, said heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene,and there bei-n g- no translational initiation codon sequences prior to said heterologous gene, 'and optionally recovering the resulting predetermined protein.
Claim 3
Dependent Claim
38 words
A method as claimed in Claim 1, wherein said heterologous gene for said predetermined protein is under the control of the B. licheniformis beta-lactamase transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site - sequences.
Claim 4
Dependent Claim
18 words
A method as claimed in Claim 1 or Claim 2, wherein said predetermined protein is a eukaryotic protein.
Claim 5
Dependent Claim
15 words
'A method as claimed in Claim 3, wherein said predetermined protein is a mammalian protein.
Claim 6
Dependent Claim
15 words
A method as claimed in Claim 4, wherein said predetermined protein is human fibroblast interferon.
Claim 7
Dependent Claim
113 words
A microorganism capable of producing a predetermined protein through expression, comprising a B. subtilis host microorganism and a plasmid therein capable of replication in B. subtilis or capable of integrating into the B. subtilis bacterial chromosome, said plasmid having a heterologous gene therein coding for a predetermined protein capable of expression in B . subtilis, said heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene, and there being no translational initiation codon prior to said heterologous gene.
Claim 8
Dependent Claim
113 words
A microorganism capable of producing a predetermined protein through expression, comprising a B. subtilis host microorganism and a plasmid therein capable of replication in B. subtilis or capable of integrating into the B. subtilis bacterial chromosome, said plasmid having a heterologous gene therein coding for a predetermined protein capable of expression in B . subtilis, said heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene, and there being no translational initiation codon prior to said heterologous gene.
Claim 9
Independent Claim
104 words
A method for creating a microorganism capable of producing a predetermined protein through expression, comprising forming a plasmid capable of replication in B . subtilis or capable of integrating into the B.subtilis bacterial chromosome, and having a heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene, and there being no translational initiation codon sequences prior to said heterologous gene, and introducing said plasmid to a B.subtilis host microorganism.
Claim 10
Independent Claim
104 words
A method for creating a microorganism capable of producing a predetermined protein through expression, comprising forming a plasmid capable of replication in B . subtilis or capable of integrating into the B.subtilis bacterial chromosome, and having a heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene, and there being no translational initiation codon sequences prior to said heterologous gene, and introducing said plasmid to a B.subtilis host microorganism.
Claim 11
Dependent Claim
92 words
A plasmid capable of replication in B. subtilis or capable of integrating into the B.subtilis bacterial chromosome, said plasmid having a heterologous gene therein coding for a predetermined protein capable of expression in B.subtilis, said heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene, and there being no translational initiation codon prior to said heterologous gene.
Claim 12
Dependent Claim
92 words
A plasmid capable of replication in B. subtilis or capable of integrating into the B.subtilis bacterial chromosome, said plasmid having a heterologous gene therein coding for a predetermined protein capable of expression in B.subtilis, said heterologous gene including a translational initiation codon sequence and being under the control of operable transcriptional and translational regulatory signals including operator, promoter and ribosomal binding site sequences, said operable transcriptional and translational regulatory signals being indigenous to a source other than said heterologous gene, and there being no translational initiation codon prior to said heterologous gene.
Claim 13
Dependent Claim
41 words
A microorganism as claimed in Claim 6, or a method as claimed in Claim 7, or a plasmid as claimed in Claim 8, and respectively further defined by the specific feature set out in any one of Claims 2 to 5.
Claim 14
Dependent Claim
48 words
lO. An intracellular protein produced by a method as claimed in any one of Claims 1 to 5 or by a microorganism as claimed in Claim 6 or Claim 9 or by a microorganism created in accordance with a method as claimed in Claim 7 or Claim 9.
Claim 15
Dependent Claim
30 words
A transformant bacterial culture cloned from one or more microorganisms as claimed in Claim 6 or Claim 9, the members of such culture being capable of expressing said predetermined protein.
References Cited (Backward Citations)
No citation records indexed.
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