EP0086537A1
Expired / Lapsed
EP
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A process for the in-vitro transformation of plant protoplasts with plasmid DNA
Assignee: Leiden University
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Filing: 1983-02-10
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Granted: —
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Expires: 2003-02-10
Abstract
Abstract Genetic properties of higher plants are transformed by incubating protoplasts of the plants together with plasmid DNA. The incubation is effected in the presence of polyethylene glycol and calcium ions. Subsequently-during after-incubation-the calcium ion concentration in the incubation medium is gradually increased and at the same time the polyethylene glycol concentration is decreased. The resulting aggregates are separated, grown further separately, and examined for modification of their genetic properties.
Inventors
- 👤 Robbert Adriaan Schilperoort
- 👤 Franciscus Andries Krens
- 👤 George Joseph Wullems
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 05:00
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/82; C12N15/8201; C12N15/8206
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
92 words
A process for transforming genetic properties of higher plants by inserting in,to the DNA of these plants the DNA of a plasmid that is foreign to these plants, or any part thereof, characterized by incubating protoplasts of higher plants together with plasmid DNA in the presence of polyethylene glycol and calcium ions, subsequently - during the after-incubation - gradually increasing both the calcium ion concentration in the incubation medium, and decreasing the polyethylene glycol concentration, separating the resulting aggregates, growing these further separately, and examining them for modification of their genetic properties.
Claim 2
Dependent Claim
92 words
A process for transforming genetic properties of higher plants by inserting in,to the DNA of these plants the DNA of a plasmid that is foreign to these plants, or any part thereof, characterized by incubating protoplasts of higher plants together with plasmid DNA in the presence of polyethylene glycol and calcium ions, subsequently - during the after-incubation - gradually increasing both the calcium ion concentration in the incubation medium, and decreasing the polyethylene glycol concentration, separating the resulting aggregates, growing these further separately, and examining them for modification of their genetic properties.
Claim 3
Dependent Claim
31 words
A process according to claim 1, characterized by selecting a polyethylene glycol concentration of 12-15% (mg/cm 3 ) and a calcium ion concentration of about 40 mmoles/1 during the first incubation.
Claim 4
Dependent Claim
36 words
A process according to claims 1-2, characterized in that during the after-incubation the calcium ion concentration is increased to 100-125 mmoles/1 with a simultaneous decrease in the polyethylene glycol concentration to about 1.6-2% (mg/cm 3 ).
Claim 5
Dependent Claim
60 words
A process according to claims 1-3, characterized in that during the after-incubation the calcium ion concentration is increased incrementally to 125 mmoles/1 with simultaneous incremental decrease of the polyethylene glycol concentration to 1.8% (mg/cm 3 ) 5. A process according to claims 1-4, characterized in that the incubation is carried out in the presence of DNA molecules as a carrier.
Claim 6
Dependent Claim
24 words
A process according to claims 1-5, characterized in that the incubation is carried out in the presence of calf thymus DNA as a carrier.
Claim 7
Dependent Claim
27 words
A process according to claims 1-6, characterized in that during the incubation the weight ratio of foreign plasmid DNA to calf thymus DNA is 1 : 5.
Claim 8
Dependent Claim
17 words
Protoplast material of higher plants with parts of a foreign plasmid DNA inserted in the DNA thereof.
Claim 9
Dependent Claim
17 words
Protoplast material of higher plants with parts of a foreign plasmid DNA inserted in the DNA thereof.
Claim 10
Dependent Claim
20 words
Mutants of higher plants with genetically modified properties, produced by the process according to one or more of claims 1-7.
Claim 11
Dependent Claim
32 words
A process for the preparation of chemical and/or pharmaceutical products, characterized by culturing cells produced by the process as claimed in one or more of claims 1-7, and isolating the desired substance.
Claim 12
Dependent Claim
20 words
A process according to claim 10, characterized in that culturing is effected by means of fermentation and possible subsequent immobilization.
References Cited (Backward Citations)
No citation records indexed.
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