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EP0138644A1 Expired / Lapsed EP ⏱️ 0.0 years remaining

Novel plasmid vectors

Assignee: Mitsubishi Chemical Corp
•
Filing: 1984-08-23
•
Granted: —
•
Expires: 2004-08-23
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Abstract

Abstract A novel plasmid vector is provided according to the present invention comprising a fragment which comprises the regulator region containing the ompF promoter of a Gram-negative bacterium, the region coding for the signal peptide of the OmpF protein and a part of the ompF structural gene. The DNA fragment is inserted into a relaxed plasmid derived from Escherichia coli. A method for producing a desired protein is a Iso provided in the invention which comprises introducing the plasmid vector of the invention comprising a gene coding for the desired protein into a host E. coli strain the protease activity of which is lost or reduced, cultivating the host and allowing the host cell to secrete the produced protein through the cytoplasmic membrane and the outer membrane into the culture medium. The protein produced in the host cell in a precursor form comprising the signal peptide is processed and secreted into the culture medium in the form of a fused protein comprising a part of the OmpF protein.

Inventors

  • 👤 Shoji Mizushima
  • 👤 Kenzo Nakamura
  • 👤 Kenji Nagahari

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-11-08 23:41
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C07; C07K; C07K14/00; C07K14/195; C07K14/24; C07K14/245; C07K14/435; C07K14/665; C07K14/67; C07K14/675; C07K2319/00; C07K2319/01; C07K2319/02

Claims Analysis

Total Claims: 7
Claim 1
Independent Claim 52 words
A plasmid vector comprising a fragment which comprises the regulator region containing the ompF promoter of a Gram-negative bacterium, the region coding for the signal peptide of the OmpF protein and 50 or less nucleotides of the ompF structural gene, the fragment being inserted into a relaxed plasmid derived from Escherichia coli.
Claim 2
Dependent Claim 52 words
A plasmid vector comprising a fragment which comprises the regulator region containing the ompF promoter of a Gram-negative bacterium, the region coding for the signal peptide of the OmpF protein and 50 or less nucleotides of the ompF structural gene, the fragment being inserted into a relaxed plasmid derived from Escherichia coli.
Claim 3
Dependent Claim 33 words
The plasmid vector in accordance with claim 1, in which the fragment is the AluI fragment, the AluI-BqlII fragment or the AluI-PstI (partial) fragment which contains the ompF promoter of a Gram-negative bacterium.
Claim 4
Dependent Claim 22 words
The plasmid vector in accordance with claim 1, in which the fragment comprises 30 or less nucleotides of the ompF structural gene.
Claim 5
Dependent Claim 23 words
The plasmid vector in accordance with claim 1, in which the fragment comprises from 20 to 26 nucleotides of the ompF structural gene.
Claim 6
Independent Claim 63 words
A method for producing a desired protein comprising introducing a plasmid vector containing a gene coding for the signal peptide of Escherichia coli and a gene coding for the desired protein into a host Escherichia coli strain, the protease activity of which is lost or reduced, cultivating the host and allowing the host cell to secrete the produced protein out of the cell.
Claim 7
Independent Claim 63 words
A method for producing a desired protein comprising introducing a plasmid vector containing a gene coding for the signal peptide of Escherichia coli and a gene coding for the desired protein into a host Escherichia coli strain, the protease activity of which is lost or reduced, cultivating the host and allowing the host cell to secrete the produced protein out of the cell.

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