EP0118393A2
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Methods and compositions for expression of competent eukaryotic gene products
Assignee: Cessione rothwell Property Ltd
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Filing: 1984-02-06
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Granted: —
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Expires: 2004-02-06
Abstract
Abstract Heat-shock gene control elements from different eukaryotic cells are utilized to provide expression of a competent gene products in the same or similar eukaryotic as well as in procaryotic host cells. The control elements can be joined to a suitable eukaryotic replication system to form an expression vector, or may be joined to the gene of interest and introduced directly into the host genome.
Inventors
- 👤 Peter Bromley
- 👤 Richard Voellmy
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 12:36
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/11; C12N15/62; C12N15/66; C12N15/70; C12N15/85; C07; C07K; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/35; C07K2319/61; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 45
Claim 1
Independent Claim
123 words
A "gene expression unit - host cell conbination system" wherein the "gene expression unit" conprises (a) an expression control region of less than 100 kb. derived from a eucaryotic heat-shock protein gene; (b) at least one site with which a structural gene of interest (c) is associated in order to come under the transcriptional and/or translational control of said expression control region (a), and wherein the "host cell system" carports cells which are either from bacterial origin or from at least one species similar to those from which the said expression control region was isolated and in which control region (a) linked functionally to the said structural gene (c) of interest is present in order to produce the product of said structural gene.
Claim 2
Dependent Claim
123 words
A "gene expression unit - host cell conbination system" wherein the "gene expression unit" conprises (a) an expression control region of less than 100 kb. derived from a eucaryotic heat-shock protein gene; (b) at least one site with which a structural gene of interest (c) is associated in order to come under the transcriptional and/or translational control of said expression control region (a), and wherein the "host cell system" carports cells which are either from bacterial origin or from at least one species similar to those from which the said expression control region was isolated and in which control region (a) linked functionally to the said structural gene (c) of interest is present in order to produce the product of said structural gene.
Claim 3
Dependent Claim
34 words
A "gene expression unit - host cell system" as in claim 1, wherein the structural gene of interest is associated by being inserted into said at least one site of the gene expression unit.
Claim 4
Dependent Claim
26 words
A "gene expression unit - host cell system" as in claim 1, wherein the heat-shock gene control region is derived from heat-shock genes from eucaryotic organisms.
Claim 5
Dependent Claim
27 words
A "gene expression unit - host cell system" as in claim 3, wherein the heat-shock gene control region is derived from the genomic DNA of Drosophila melanogaster.
Claim 6
Dependent Claim
35 words
A "gene expression unit - host cell system" as in claim 4, wherein the heat-shock gene control region is derived from any one of the genes encoding the 70 kilo-dalton heat-shock proteins of Drosophila melanogaster.
Claim 7
Dependent Claim
27 words
A "gene expression unit - host cell system" as in claim 1, wherein at least part of said functional heat-shock gene control region is of synthetic origin.
Claim 8
Dependent Claim
36 words
A "gene expression unit - host cell system" as in claim 1, in which said expression unit further comprises at least one selectable marker linked to the expression unit which allows for selection of transformed hosts.
Claim 9
Dependent Claim
37 words
A "gene expression unit - host cell system" as in claim 1, in which said expression unit further comprises a procaryotic replication system which allows propagation of the expression unit, and at least one antibiotic resistance gene.
Claim 10
Dependent Claim
27 words
A "gene expression unit - host cell system" as in claim 1, wherein said expression unit is linked to a fragment containing a eucaryotic extrachomosomal replication system.
Claim 11
Dependent Claim
39 words
A "gene expression unit - host cell system" as in claim 1 wherein the eucaryotic heat-shock gene control region is linked to a cellular or viral transcription enhancer element to allow for constitutive expression of the gene of interest.
Claim 12
Dependent Claim
31 words
A "gene expression unit - host cell system" as in claim 1, wherein a heat-shock promoter element is linked to the complete RNA coding region of a structural gene of interest.
Claim 13
Dependent Claim
32 words
A "gene expression unit - host cell system" as in claim 1, wherein a eucaryotic heat-shock gene promoter-RNA leader segment is linked to the protein-coding region of a structural gene of interest.
Claim 14
Dependent Claim
35 words
A "gene expression unit - host cell system" as in claim 1, wherein a eucaryotic heat-shock gene control element is linked to at least part of the DNA-coding region of a structural gene of interest.
Claim 15
Dependent Claim
8 words
Plasmid pRVl5 as a new conposition of matter.
Claim 16
Dependent Claim
8 words
Plasmid pRVl5 as a new conposition of matter.
Claim 17
Dependent Claim
8 words
Plasmid PR84 as a new conposition of matter.
Claim 18
Dependent Claim
8 words
Plasmid PR84 as a new conposition of matter.
Claim 19
Dependent Claim
28 words
A method for preparing the gene expression - host cell combination of claim 1, comprising introducing the expression unit into host cells by cotransformation with a selectable marker.
Claim 20
Dependent Claim
19 words
A method as in claim 16, wherein the expression unit is introduced into host cells with an anplifiable gene.
Claim 21
Dependent Claim
19 words
A method as in claim 16, wherein the expression unit is introduced into host cells by transformation or transfection.
Claim 22
Dependent Claim
34 words
An expression vector corrprising: a eukaryotic extrachromosomal replication system; an expression control region derived from a eukaryotic heat-shock protein; and at least one insertion site under the transcriptional control of the expression control region.
Claim 23
Dependent Claim
34 words
An expression vector corrprising: a eukaryotic extrachromosomal replication system; an expression control region derived from a eukaryotic heat-shock protein; and at least one insertion site under the transcriptional control of the expression control region.
Claim 24
Dependent Claim
25 words
An expression vector as in claim 19, said vector being free of a translational start codon between the expression control region and the insertion site.
Claim 25
Dependent Claim
17 words
An expression vector as in claim 19, having a translational start codon upstream from the insertion site.
Claim 26
Dependent Claim
22 words
An expression vector as in claim 19, wherein the heat-shock gene control region is derived from the genomic DNA of Drosophila melanogaster.
Claim 27
Dependent Claim
29 words
An expression vector as in claim 22, wherein the heat-shock gene control region is derived from the heat-shock gene which encodes the 70 kilodalton heat-shock protein of Drosophila melanogaster.
Claim 28
Dependent Claim
20 words
An expression vector as in claim 19, further comprising a prokaryotic replication system which allows stable maintenance in prokaryotic hosts.
Claim 29
Dependent Claim
21 words
An expression vector as in claim 19, further comprising at least one selectable marker which allows for selection of transformed hosts.
Claim 30
Dependent Claim
51 words
An expression vector ccnprising: a eukaryotic extrachromosomal replication system; a heat-shock gene control region substantially homologous to a 650 base pair sequence which controls transcription of the 70 kilo- dalton heat-shock protein in Drosophila melanogaster; and at least one insertion site under the transcriptional control of the heat-shock gene control region.
Claim 31
Dependent Claim
51 words
An expression vector ccnprising: a eukaryotic extrachromosomal replication system; a heat-shock gene control region substantially homologous to a 650 base pair sequence which controls transcription of the 70 kilo- dalton heat-shock protein in Drosophila melanogaster; and at least one insertion site under the transcriptional control of the heat-shock gene control region.
Claim 32
Dependent Claim
36 words
A DNA construct of less than 15 kbp which includes an expression control region derived from a .eukaryotic heat-shock gene and at least one insertion site under the transcriptional control of the heat-shock gene control region.
Claim 33
Dependent Claim
36 words
A DNA construct of less than 15 kbp which includes an expression control region derived from a .eukaryotic heat-shock gene and at least one insertion site under the transcriptional control of the heat-shock gene control region.
Claim 34
Dependent Claim
21 words
A DNA construct as in claim 27, wherein the heat-shock gene which encodes the 70 kilodalton heat-shock protein of Drosophila melanogaster.
Claim 35
Independent Claim
52 words
A method for producing competent gene products, said method comprising: joining a structural gene to an expression control region derived from a eukaryotic heat-shock gene; introducing said structural gene and control region into a eukaryotic host suitable for expression of the structural gene; and growing said host, whereby said product is produced.
Claim 36
Independent Claim
52 words
A method for producing competent gene products, said method comprising: joining a structural gene to an expression control region derived from a eukaryotic heat-shock gene; introducing said structural gene and control region into a eukaryotic host suitable for expression of the structural gene; and growing said host, whereby said product is produced.
Claim 37
Dependent Claim
26 words
A method as in claim 29, wherein the structural gene and control region are joined together in an expression vector having an eukaryotic extrachromosomal replication system.
Claim 38
Dependent Claim
24 words
A method as in claim 29, wherein the structural gene and control region are introduced to the host by co-transformation with a selectable marker.
Claim 39
Dependent Claim
27 words
A method as in claim 29, wherein the expression control region is derived from a heat-shock gene which encodes a 70 kilodalton heat-shock protein in Drosophila melanogaster.
Claim 40
Dependent Claim
20 words
A method as in claim 29, wherein the heat-shock gene is derived from a different eukaryotic species from the host.
Claim 41
Dependent Claim
14 words
A method as in claim 29, wherein the structural gene is a mammalian gene.
Claim 42
Dependent Claim
14 words
A method as in claim 34, wherein the host is a mammalian cell culture.
Claim 43
Dependent Claim
14 words
A method as in claim 29, wherein the structural gene is a human gene..
Claim 44
Dependent Claim
14 words
A method as in claim 36, wherein the host is a mammalian cell culture.
Claim 45
Dependent Claim
15 words
A method as in claim 37, wherein the host is a COS I cell culture.
References Cited (Backward Citations)
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