Patent EP0035831A2 successfully refreshed from Google Patents.
EP0035831A2
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Method for making genetically modified microorganisms
Assignee: Imperial Chemical Industries Ltd
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Filing: 1981-02-10
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Granted: —
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Expires: 2001-02-10
Abstract
Abstract A method for making a genetically modified microorganism wherein DNA material, particularly material comprising a gdh gene, is transferred from a donor microorganism to a recipient microorganism using a plasmid vector. The invention relates in particular to the use of multicopy plasmids and/or DNA fragments having at their ends target sites for Sall restriction enzymes. The method may involve a plurality of steps, transfer of DNA fragments into intermediate microorganisms and use of a plurality of plasmids. Genetically modified microorganisms and a process for the production of single cell protein using the genetically modified microorganisms are also claimed.
Inventors
- 👤 Michael John Worsey
- 👤 John David Windass
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 06:44
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0012; C12N9/0014; C12N9/0016; C12N1/00; C12N1/20; C12N1/205; C12N1/32; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/78; C12R; C12R2001/00; C12R2001/01
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
79 words
1 . A method for making a genetically modified microorganism by incorporating DNA material comprising a gdh gene into a microorganism deficient in this gene, so that expression of the gdh gene product, i.e. the active GDH enzyme, occurs in the previously gdh gene-deficient microorganism, which method comprises the steps of (a) introducing into a first microorganism having the gdh gene a plasmid vector for said gene, and (b) transferring said gene on said vector into a second microorganism.
Claim 2
Dependent Claim
79 words
1 . A method for making a genetically modified microorganism by incorporating DNA material comprising a gdh gene into a microorganism deficient in this gene, so that expression of the gdh gene product, i.e. the active GDH enzyme, occurs in the previously gdh gene-deficient microorganism, which method comprises the steps of (a) introducing into a first microorganism having the gdh gene a plasmid vector for said gene, and (b) transferring said gene on said vector into a second microorganism.
Claim 3
Independent Claim
51 words
A method for making a genetically modified microorganism by transferring DNA material comprising one or more genes from a donor microorganism having said gene or genes to a recipient microorganism deficient therein wherein the DNA material is transferred into the recipient microorganism on a plasmid vector formed from a multicopy plasmid.
Claim 4
Independent Claim
51 words
A method for making a genetically modified microorganism by transferring DNA material comprising one or more genes from a donor microorganism having said gene or genes to a recipient microorganism deficient therein wherein the DNA material is transferred into the recipient microorganism on a plasmid vector formed from a multicopy plasmid.
Claim 5
Independent Claim
57 words
A method for making a genetically modified microorganism by transferring DNA material comprising the gdh gene from a donor microorganism having said gene to a recipient microorganism deficient therein which comprises a step wherein the gdh gene containing DNA material is transferred on a DNA fragment having at its ends target sites for the SalI restriction enzyme.
Claim 6
Independent Claim
57 words
A method for making a genetically modified microorganism by transferring DNA material comprising the gdh gene from a donor microorganism having said gene to a recipient microorganism deficient therein which comprises a step wherein the gdh gene containing DNA material is transferred on a DNA fragment having at its ends target sites for the SalI restriction enzyme.
Claim 7
Dependent Claim
34 words
A method according to claim 1 wherein the second microorganism is an intermediate and the gdh gene is transferred via one or more intermediate microorganisms from the first microorganism to the gdh gene-deficient microorganism.
Claim 8
Dependent Claim
31 words
A method according to claim 2 which comprises a plurality of steps involving at least one intermediary microorganism during transfer of the DNA material from the donor to the recipient microorganism.
Claim 9
Dependent Claim
17 words
A method according to claim 2 or claim 5 wherein the gene transferred is a gdh gene.
Claim 10
Dependent Claim
22 words
A method according to claim 3 wherein the DNA fragment transferred is transferred on a plasmid vector formed from a multicopy plasmid.
Claim 11
Independent Claim
23 words
A method according to any one of the preceding claims wherein a gdh gene is transferred into a gram-negative bacterium lacking this gene.
Claim 12
Dependent Claim
18 words
A method according to claim 8 wherein the gram-negative bacterium is a strain of the species Methylophilus methylotrophus.
Claim 13
Dependent Claim
15 words
A method according to claim 9 wherein the strain is Methylophilus methylotrophus NCIB No. 10515.
Claim 14
Independent Claim
28 words
A method according to any one of the preceding claims which comprises a step in which the git gene product is inactivated by mutation in gdh gene-deficient microorganism.
Claim 15
Independent Claim
25 words
A method according to any one of the preceding claims- wherein the gdh gene containing DNA material is obtained from a strain of E. coli.
Claim 16
Independent Claim
41 words
A method according to any one of the preceding claims wherein the gdh gene containing DNA material is transferred into the gdh gene deficient microorganism on a plasmid vector formed from an Inc Q, Inc P or Inc W group plasmid.
Claim 17
Independent Claim
17 words
A method for making a genetically modified microorganism substantially as described and as shown in Example 1.
Claim 18
Independent Claim
17 words
A method for making a genetically modified microorganism substantially as described and as shown in Example 1.
Claim 19
Dependent Claim
24 words
Genetically modified microorganisms produced by a method according to any one of the preceding claims and variants and mutants of such genetically modified microorganisms.
Claim 20
Dependent Claim
15 words
Genetically modified microorganisms according to claim 15 which are genetically modified strains of gram-negative bacteria.
Claim 21
Dependent Claim
26 words
Genetically modified microorganisms according to claim 16 which are strains of the species Methylophilus methylotrophus genetically modified by incorporation of DNA material comprising a gdh gene.
Claim 22
Dependent Claim
13 words
The microorganism Methylophilus methylotrophus strain NCIB No. 11585 and variants and mutants thereof.
Claim 23
Dependent Claim
13 words
The microorganism Methylophilus methylotrophus strain NCIB No. 11585 and variants and mutants thereof.
Claim 24
Dependent Claim
93 words
19 A process for the production of single cell protein and/ or an amino acid wherein a gram-negative bacterium genetically modified by incorporation of DNA material comprising a gdh gene and produced by a method according to any one of claims 1 to 14 is aerobically cultured in an aqueous medium containing methanol as a source of assimilable carbon and ammonia and/or ammonium ion at a concentration sufficient to maintain the GDH reaction mechanism and the single cell protein, a composition containing it and/or an amino acid is recovered from the culture medium.
Claim 25
Dependent Claim
21 words
A process according to claim 19 wherein the genetically modified microorganism is a genetically modified strain of the species Methylophilus methylotrophus.
Claim 26
Dependent Claim
24 words
A process according to claim 20 wherein the genetically modified microorganism is Methylophilus methylotrophus strain NCIB No. 11585 or a variant or mutant thereof.
Claim 27
Dependent Claim
13 words
Dead cells of microorganisms according to any one of claims 15 to 17.
Claim 28
Dependent Claim
13 words
Protein derived from microorganisms according to any one of claims 15 to 17.
Claim 29
Dependent Claim
20 words
A foodstuff for animal or human consumption containing dead cells according to claim 22 or protein according to claim 23.
Claim 30
Dependent Claim
18 words
Modified plasmids produced at intermediate stages of a method according to any one of claims 1 to 14.
Claim 31
Dependent Claim
9 words
Modified R300B, pRP301, pACYC184 plasmids incorporating the gdh gene.
Claim 32
Dependent Claim
9 words
Modified R300B, pRP301, pACYC184 plasmids incorporating the gdh gene.
References Cited (Backward Citations)
No citation records indexed.
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