Patent EP0123228A2 successfully refreshed from Google Patents.
EP0123228A2
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Hybrid DNA synthesis of mature insulin-like growth factors
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1984-04-13
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Granted: —
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Expires: 2004-04-13
Abstract
Abstract Methods and compositions are provided for efficient production of human insulin-like growth factor. Synthetic IGF I and IGF II genes are joined to leader and processing signals which provide for expression and secretion of the gene product in yeast. Enhanced yields of the product may then be recovered from the nutrient medium. Yeast strains S. cerevisiae AB103 (pYIGF-I-10/1) and AB103 (pYIGF-Il-10/1) were deposited at the American Type Culture Collection on April 23, 1983 and granted Accession Nos. 20673 and 20674, respectively.
Inventors
- 👤 Philip J. Barr
- 👤 James P. Merryweather
- 👤 Guy Mullenbach
- 👤 Mickey S. Urdea
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 11:11
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/65; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/81; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/036; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
79 words
A method for efficient production of a human insulin-like growth factor (IGF) in a suitable host, said method comprising growing host cells containing a functional DNA construct having the gene encoding said IGF joined in proper reading frame with secretory leader and processing signal sequences recognized by said host to form a structural gene downstream from and under the transcriptional regulatory control of a transcriptional initiation region in a vector compatible with said host, and isolating secreted human IGF.
Claim 2
Independent Claim
79 words
A method for efficient production of a human insulin-like growth factor (IGF) in a suitable host, said method comprising growing host cells containing a functional DNA construct having the gene encoding said IGF joined in proper reading frame with secretory leader and processing signal sequences recognized by said host to form a structural gene downstream from and under the transcriptional regulatory control of a transcriptional initiation region in a vector compatible with said host, and isolating secreted human IGF.
Claim 3
Dependent Claim
21 words
A method as in claim 1, wherein the IGF gene is a synthetic gene having codons preferentially utilized by the host.
Claim 4
Dependent Claim
22 words
A method as in claim 1, wherein the human IGF gene is a synthetic IGF I gene having the following nucleotide sequence:
Claim 5
Dependent Claim
22 words
A method as in claim 1, wherein the human IGF gene is a synthetic IGF II gene having the following nucleotide sequence:
Claim 6
Dependent Claim
12 words
A method as in claim 1, wherein the host is a yeast.
Claim 7
Dependent Claim
22 words
A method as in claim 1, wherein the host is a yeast and the vehicle is a derivative of the 2pm plasmid.
Claim 8
Independent Claim
192 words
A method for expressing human insulin-like growth factor (IGF) in yeast resulting in secretion of said IGF promoted by yeast secretion and processing signals, said method comprising: preparing a first DNA fragment containing a first DNA sequence encoding for IGF and having the 5'-terminus of the coding strand at or downstream from the first base of said DNA sequence; preparing a second DNA fragment containing a second DNA sequence encoding for yeast-recognizable secretion and processing signals and having the 3'- terminus of the coding strand at or upstream from the terminal base of said processing signal, wherein at least one of said first and second fragments has its terminus internal to the coding sequence resulting in missing base pairs; joining said first and second DNA fragments by means of an adaptor encoding for the missing base pairs of said first and second DNA fragments, wherein said first and second DNA fragments are in the same reading frame; providing a termination codon downstream of and adjacent to the first fragment to provide a "pre"-IGF gene; and cloning said "pre"-IGF gene in an expression vector in yeast, wherein said "pre"-IGF is secreted and processed.
Claim 9
Independent Claim
192 words
A method for expressing human insulin-like growth factor (IGF) in yeast resulting in secretion of said IGF promoted by yeast secretion and processing signals, said method comprising: preparing a first DNA fragment containing a first DNA sequence encoding for IGF and having the 5'-terminus of the coding strand at or downstream from the first base of said DNA sequence; preparing a second DNA fragment containing a second DNA sequence encoding for yeast-recognizable secretion and processing signals and having the 3'- terminus of the coding strand at or upstream from the terminal base of said processing signal, wherein at least one of said first and second fragments has its terminus internal to the coding sequence resulting in missing base pairs; joining said first and second DNA fragments by means of an adaptor encoding for the missing base pairs of said first and second DNA fragments, wherein said first and second DNA fragments are in the same reading frame; providing a termination codon downstream of and adjacent to the first fragment to provide a "pre"-IGF gene; and cloning said "pre"-IGF gene in an expression vector in yeast, wherein said "pre"-IGF is secreted and processed.
Claim 10
Dependent Claim
12 words
A method according to claim 7, wherein said IGF is IGF I.
Claim 11
Dependent Claim
12 words
A method according to claim 7, wherein said IGF is IGF II.
Claim 12
Dependent Claim
17 words
A method according to claim 7, wherein said expression vector includes a replication system recognized by bacteria.
Claim 13
Dependent Claim
18 words
A method according to claim 7, wherein said yeast replication system comprises the 2pm plasmid or portion thereof.
Claim 14
Dependent Claim
23 words
A DNA construct comprising a coding sequence coding for human insulin-like growth factor (IGF) under transcriptional regulation of regulatory signals utilized by yeast.
Claim 15
Dependent Claim
23 words
A DNA construct comprising a coding sequence coding for human insulin-like growth factor (IGF) under transcriptional regulation of regulatory signals utilized by yeast.
Claim 16
Dependent Claim
16 words
A DNA construct according to claim 12, wherein said IGF coding sequence codes for IGF I.
Claim 17
Dependent Claim
16 words
A DNA construct according to claim 12, wherein said IGF coding sequence codes for IGF II.
Claim 18
Dependent Claim
14 words
A DNA construct according to claim 12 including a replication system recognized by yeast.
Claim 19
Dependent Claim
27 words
A DNA construct according to claim 12, where at least a portion of said coding sequence has codons preferentially utilized by yeast and different from human codons.
Claim 20
Dependent Claim
17 words
A DNA construct according to claim 12, wherein said construct includes a replication system utilized by bacteria.
Claim 21
Dependent Claim
19 words
A DNA construct according to claim 15 wherein said yeast replication system comprises the 2pm plasmid or portion thereof.
Claim 22
Dependent Claim
10 words
A DNA construct according to claim 12, including the sequence:
Claim 23
Dependent Claim
10 words
A DNA construct according to claim 12, including the sequence:
Claim 24
Dependent Claim
27 words
A DNA construct comprising a sequence coding for human insulin-like growth factor joined to transcriptional and translational regulatory sequences for expression of a cellular host in culture.
Claim 25
Dependent Claim
27 words
A DNA construct comprising a sequence coding for human insulin-like growth factor joined to transcriptional and translational regulatory sequences for expression of a cellular host in culture.
References Cited (Backward Citations)
No citation records indexed.
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