EP0251744A2
Expired / Lapsed
EP
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Process for the genetic modification of yeast
Assignee: Albumedix Ltd
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Filing: 1987-06-26
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Granted: —
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Expires: 2007-06-26
Abstract
Abstract Yeast is genetically modified by transformation with an integration vector comprising two copies of a homologous 2 µm plasmid DNA sequence in direct orientation relative to one another and encompassing the said DNA sequence, and then isolating, from the transformed yeast obtained, cells containing the endogenous 2 µm plasmid modified by incorporation of the said DNA sequence but not containing the said vector. The resulting yeast can be maintained under non-selective growth conditions.
Inventors
- 👤 Edward Hinchliffe
- 👤 Christine Jane Fleming
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 07:09
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/76; C07K14/765; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/81
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
89 words
Process for the genetic modification of yeast by incorporation into the endogenous 2 µm plasmid of a DNA sequence coding for a protein or peptide of interest which comprises first transforming yeast with an integration vector comprising two copies of a homologous 2 µm plasmid DNA sequence in direct orientation relative to one another and encompassing the said DNA sequence, and then isolating, from the transformed yeast obtained, cells containing the endogenous 2 µm plasmid modified by incorporation of the said DNA sequence but not containing the said vector.
Claim 2
Dependent Claim
89 words
Process for the genetic modification of yeast by incorporation into the endogenous 2 µm plasmid of a DNA sequence coding for a protein or peptide of interest which comprises first transforming yeast with an integration vector comprising two copies of a homologous 2 µm plasmid DNA sequence in direct orientation relative to one another and encompassing the said DNA sequence, and then isolating, from the transformed yeast obtained, cells containing the endogenous 2 µm plasmid modified by incorporation of the said DNA sequence but not containing the said vector.
Claim 3
Dependent Claim
37 words
Process according to claim 1 in which the integration vector also comprises an extraneous DNA sequence separated from the DNA sequence coding for the protein or peptide of interest by the said homologous sequences in direct orientation.
Claim 4
Dependent Claim
27 words
Process according to claim 2 in which the said extraneous DNA sequence is a DNA sequence heterologous to yeast which assists propagation of the vector in bacteria.
Claim 5
Dependent Claim
45 words
Process according to any of claims 1 to 3 in which the said vector also includes a selectable marker DNA sequence which is not separated from the DNA sequence which codes for the protein or peptide of interest by the homologous sequences in direct orientation.
Claim 6
Dependent Claim
20 words
Process according to claim 4 in which the selectable marker DNA sequence is a gene coding for resistance to copper.
Claim 7
Dependent Claim
28 words
Process according to any one of claims 1 to 5 in which the said vector includes an origin of replication endogenous to the 2 µm plasmid of yeast.
Claim 8
Dependent Claim
32 words
Process according to any one of claims 1 to 6 in which the DNA sequence coding for a protein or peptide of interest codes for human serum albumin or a derivative thereof.
Claim 9
Dependent Claim
48 words
Process according to any one of claims 1 to 7 in which each of the homologous 2 µm plasmid sequences in direct orientation comprises a 703 base pair region of DNA from the endogenous 2 µm plasmid of yeast bounded by an EcoRI site and a XbaI site.
Claim 10
Dependent Claim
17 words
Process according to any one of claims 1 to 8 in which the yeast is brewing yeast.
Claim 11
Dependent Claim
61 words
A 2 µm plasmid vector comprising two copies of a homologous 2 µm plasmid DNA sequence in direct orientation, a DNA sequence assisting propagation of the plasmid outside yeast, and a DNA sequence coding for a heterologous protein or peptide of interest separated from the said DNA sequence assisting propagation of the plasmid by the said homologous sequences in direct orientation.
Claim 12
Dependent Claim
61 words
A 2 µm plasmid vector comprising two copies of a homologous 2 µm plasmid DNA sequence in direct orientation, a DNA sequence assisting propagation of the plasmid outside yeast, and a DNA sequence coding for a heterologous protein or peptide of interest separated from the said DNA sequence assisting propagation of the plasmid by the said homologous sequences in direct orientation.
Claim 13
Dependent Claim
42 words
A 2 µm plasmid vector according to claim 10 in which also includes a selectable marker DNA sequence which is not separated from the DNA sequence which codes for the protein or peptide of interest by the homologous sequences in direct orientation.
Claim 14
Dependent Claim
24 words
A 2 µm plasmid vector according to claim 11 in which the selectable marker DNA sequence is a gene coding for resistance to copper.
Claim 15
Dependent Claim
28 words
A 2 µm plasmid vector according to any one of claims 10 to 12 which includes an origin of replication endogenous to the 2 µm plasmid of yeast.
Claim 16
Dependent Claim
31 words
A 2 µm plasmid vector according to any one of claims 10 to 13 in which the heterologous protein or peptide of interest is human serum albumin or a derivative thereof.
Claim 17
Dependent Claim
52 words
A 2 µm plasmid vector according to any one of claims 10 to 14 in which each of the homologous 2 µm plasmid sequences in direct orientation comprises a 703 base pair region of DNA from the endogenous 2 µm plasmid of yeast bounded by an EcoRI site and a XbaI site.
References Cited (Backward Citations)
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