EP0068701A2
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EP
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Proinsulin genes with modified C-chain
Assignee: Canadian Patents and Development Ltd
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Filing: 1982-06-14
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Granted: —
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Expires: 2002-06-14
Abstract
Abstract Human-like proinsulin gene analogs have been synthesized by a combination of chemical and enzymatic methods. A number of different human-like proinsulin gene analogs with altered C-chains have been designed and can be readily constructed as described. As a part of the strategy, an adaptor for trimming DNA has been used to recover the A-chain insulin gene with the desired sequence from a hybrid plasmid; a related adaptor for trimming DNA has been used to shorten the C-chain gene. The synthetic proinsulin gene has been joined to a replicable cloning vehicle and the hybrid DNA transferred to a host cell. The transformed host cells can be grown under selected conditions to yield the proinsulin analogs.
Inventors
- 👤 Saran A. Narang
- 👤 Ray J. Wu
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-05 22:38
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/66; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/62
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
21 words
An insulin C-chain gene analog shortened or lengthened from the normal gene length by the deletion or addition of nucleotides, respectively.
Claim 2
Dependent Claim
21 words
An insulin C-chain gene analog shortened or lengthened from the normal gene length by the deletion or addition of nucleotides, respectively.
Claim 3
Dependent Claim
22 words
The insulin C-chain gene analog of claim 1 shortened or lengthened by the deletion or addition of nucleotides in multiples of three.
Claim 4
Dependent Claim
22 words
The insulin C-chain gene of claim I wherein the normal gene length is the duplex DNA nucleotides 91-195 of formula 2 herein.
Claim 5
Dependent Claim
35 words
An insulin C-chain partial gene as in claim 3 having the duplex DNA sequence C-1 to C-7 plus C-9 to C-16 of formula 2, suitable for ligation to appropriately adapted insulin A-chain and B-chain genes.
Claim 6
Dependent Claim
18 words
The insulin C-chain gene of claim 3 shortened 12 base pairs by deletion of fragments C-5 and C-13.
Claim 7
Dependent Claim
20 words
The insulin C-chain gene of claim 3 shortened 21 base pairs by deletion of fragments C-3, C-4, C-11 and C-12.
Claim 8
Dependent Claim
22 words
The insulin C-chain gene of claim 3 shortened 33 base pairs by deletion of fragments C-3, C-4, C-5, C-11, C-12 and C-13.
Claim 9
Dependent Claim
26 words
The insulin C-chain gene of claim 3 shortened by one of 12, 18, 24, 30, or more base pairs leftward from nucleotide 182 of Formula 2.
Claim 10
Dependent Claim
32 words
The insulin C-chain gene of claim 3 shortened by one of 6, 9, 12, 15, .... up to 81 base pairs, in multiples of 3, leftward from nucleotide 182 of Formula 2.
Claim 11
Dependent Claim
31 words
The insulin C-chain gene of claim 3 lengthened by multiples of three base pairs by ligating DNA fragments of selected lengths at the Sau3A site (5' GATC) of the C-chain gene.
Claim 12
Dependent Claim
20 words
The insulin C-chain gene analog having the formula: CGT CGT AAG CfT AAG CGT GCA GCA TTG GAA TTC GCA
Claim 13
Dependent Claim
20 words
The insulin C-chain gene analog having the formula: CGT CGT AAG CfT AAG CGT GCA GCA TTG GAA TTC GCA
Claim 14
Dependent Claim
17 words
An insulin C-chain gene including inserted nucleotides selected to form at least one restriction enzyme recognition site.
Claim 15
Dependent Claim
17 words
An insulin C-chain gene including inserted nucleotides selected to form at least one restriction enzyme recognition site.
Claim 16
Dependent Claim
26 words
Proinsulin gene analogs in which the normal C-chain DNA has been shortened by deletion of nucleotides or lengthened by addition of nucleotides in multiples of three.
Claim 17
Dependent Claim
26 words
Proinsulin gene analogs in which the normal C-chain DNA has been shortened by deletion of nucleotides or lengthened by addition of nucleotides in multiples of three.
Claim 18
Dependent Claim
13 words
The proinsulin gene analogs of claim 13 ligated to a replicable cloning vehicle.
Claim 19
Independent Claim
104 words
A method of assembling a modified proinsulin gene comprising: (a) providing a combined insulin B + partial C chain gene analog having the enzyme recognition site for.Sau3A (3' CTAG 5') in the lower DNA strand at the free partial C-chain end, (b) providing an insulin A-chain gene carrying the other part of the C-chain gene analog and having the complementary recognition site for said enzyme 5' GATC 3' at the upstream end in the upper DNA strand of the remainder of the C-chain gene analog, and (c) ligating the two genes (a) and (b) at their Sau3A sites to form the complete proinsulin gene.
Claim 20
Independent Claim
104 words
A method of assembling a modified proinsulin gene comprising: (a) providing a combined insulin B + partial C chain gene analog having the enzyme recognition site for.Sau3A (3' CTAG 5') in the lower DNA strand at the free partial C-chain end, (b) providing an insulin A-chain gene carrying the other part of the C-chain gene analog and having the complementary recognition site for said enzyme 5' GATC 3' at the upstream end in the upper DNA strand of the remainder of the C-chain gene analog, and (c) ligating the two genes (a) and (b) at their Sau3A sites to form the complete proinsulin gene.
References Cited (Backward Citations)
No citation records indexed.
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