US4830962A
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Recombinant diphtheria toxin fragments
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1984-02-09
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Granted: 1989-05-16
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Expires: 2004-02-09
Abstract
Abstract Recombinant vectors which are effective in expressing DNA sequences encoding specific fragments of diphtheria toxins at high levels in recombinant host cells are disclosed. Both a fragment consisting of the enzymatically active A chain of diphtheria toxin and a fragment consisting of both the A portion and a B portion partial sequence are constructed by use of this recombinant vector.
Inventors
- 👤 David H. Gelfand
- 👤 Lawrence I. Greenfield
- 👤 Frances C. Lawyer
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 17:22
CPC Classifications
C; C07; C07K; C07K7/00; C07K7/04; C07K7/08; A; A61; A61K; A61K39/00; A61K39/395; A61K39/44; A61K47/00; A61K47/50; A61K47/51; A61K47/68; A61K47/6801; A61K47/6803; A61K47/6811; A61K47/6817; A61K47/6835; A61K47/6851; A61K47/6855; A61K47/6889; C07K14/00; C07K14/195; C07K14/34; C07K14/415; C07K16/00; C07K16/18; C07K16/28; C07K16/30; C07K16/3015; C07K17/00; C07K17/02; C07K17/06; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/62; C12N15/63; C12N15/67; C12N15/69; C12N15/70; A61K38/00; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/33; C07K2319/55; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
94 words
An expression vecot effective in expressing a DNA sequence encoding diphtheria toxin (DT) fragment, which comprises a first DNA sequnce encoding a DT-A fragment having at its N-terminal a methioine residiue, said DNA sequence encoding a translation termination signal immediately downstream and adjacent to the C-terminal thereof, said first DNA seqeunce operably linked to a second DNA comprising a control sequence other than the native DT control sequence compatible with a transformant host cell and which does not contain between the first and second DNA sequence the native DT control sequence or portions thereof.
Claim 2
Dependent Claim
94 words
An expression vecot effective in expressing a DNA sequence encoding diphtheria toxin (DT) fragment, which comprises a first DNA sequnce encoding a DT-A fragment having at its N-terminal a methioine residiue, said DNA sequence encoding a translation termination signal immediately downstream and adjacent to the C-terminal thereof, said first DNA seqeunce operably linked to a second DNA comprising a control sequence other than the native DT control sequence compatible with a transformant host cell and which does not contain between the first and second DNA sequence the native DT control sequence or portions thereof.
Claim 3
Dependent Claim
27 words
The vector of claim 1 which further does not contain between the first and second DNA sequence the coding sequence, or portion thereof, for native DT leader.
Claim 4
Dependent Claim
29 words
The vector of claim 1 wherein said first DNA encodes a DT-A fragment which has the N-terminal sequence met-gly-ala-asp-asp and comprises an ATG encoding the N-terminal methionine residue thereof.
Claim 5
Dependent Claim
21 words
The expression vector of claim 1 wherein the DNA sequence encodes a DT-A fragment having the sequence Arg-Ser-Val-Gly-Ser-Ser at its C-terminus.
Claim 6
Dependent Claim
20 words
The vector of claim 3 wherein the DNA sequence encodes a DT-A fragment having the sequence Arg-Ser-VAl-Gly-Ser-Ser at its C-terminus.
Claim 7
Dependent Claim
18 words
The vector of claim 1 wherein the second DNA seqeence comprising a control sequence comprises the trip promoter.
Claim 8
Dependent Claim
19 words
The vector of claim 1 wherein the second DNA sequence comprises a control sequence comprsies the P L promoter.
Claim 9
Dependent Claim
8 words
The vector of claim 1 which is pTrpSmlMbo.
Claim 10
Dependent Claim
13 words
The vector of claim 1 which is P L P trp Switch A.
Claim 11
Dependent Claim
10 words
A recombinant cell transformed with the vecor of claim 1.
Claim 12
Dependent Claim
23 words
A method of producing DT-A fragment which method comprises (a) culturing the recombinant cells of claim 10, and (b) recovering the DT-A fragment.
Claim 13
Dependent Claim
95 words
An expression vector effective in expressing a DNA seqeunce encoding diphtheria toxin (DT) AB' (DT-A-B') which comprises a first DNA sequence encoding a DT-A-B' fragment having at its N-terminal a mothionine residue, said DNA seqeunce encoding a translation termination signal immediately downstream and adjacent to the C-terminal thereof, said first DNA sequence operably linked to a second DNA comprising a control sequence other than the native DT control sequence compatible with a transformant host cell and which does not contain between the first and second DNA sequence the native DT control sequence or portions thereof.
Claim 14
Dependent Claim
95 words
An expression vector effective in expressing a DNA seqeunce encoding diphtheria toxin (DT) AB' (DT-A-B') which comprises a first DNA sequence encoding a DT-A-B' fragment having at its N-terminal a mothionine residue, said DNA seqeunce encoding a translation termination signal immediately downstream and adjacent to the C-terminal thereof, said first DNA sequence operably linked to a second DNA comprising a control sequence other than the native DT control sequence compatible with a transformant host cell and which does not contain between the first and second DNA sequence the native DT control sequence or portions thereof.
Claim 15
Dependent Claim
27 words
The vector of claim 12 which further does not contain between the first and second DNA sequence the coding sequence, or portion thereof, for native DT leader.
Claim 16
Dependent Claim
29 words
The vector of claim 12 wherein said first DNA encodes a DT-A-B' fragment which has the N-terminal sequence met-gly-ala-asp-asp and comprises an ATG encoding the N-terminal methionine residue thereof.
Claim 17
Dependent Claim
22 words
The expression vector of claim 2 wherein the first DNA sequence encodes a DT-A-B' fragment having the sequence Tyr-Ser-Pro-Gly at its C-terminus.
Claim 18
Dependent Claim
21 words
The vector of claim 13 wherein the first DNA sequence encodes DT-A-B' fragment which has the sequence Tyr-Ser-Pro-Gly at its C-terminus.
Claim 19
Dependent Claim
22 words
The expression vector of claim 12 wherein the first DNA sequence encodes a DT-A-B fragment having the fragment Tyr-Ser-Pro-Gly-Cys at its C-terminus.
Claim 20
Dependent Claim
21 words
The vector of claim 13 wherein the first DNA sequence encodes a DT-A-B' fragment having the fragment Tyr-Ser-Pro-Gly-Cys at its C-terminus.
Claim 21
Dependent Claim
14 words
The vector of claim 12 wherein the second DNA sequence comprises the trp promoter.
Claim 22
Dependent Claim
15 words
The eector of claim 12 wherein the second DNA sequence comprises the P L promoter.
Claim 23
Dependent Claim
20 words
The vector of claim 20 which is selected from the group consisting of pP L OPMspRT and pP L MspRT.
Claim 24
Dependent Claim
20 words
The vector of claim 20 which is selected from the group consisting of pP L OPMspCys and pP L MspCys.
Claim 25
Dependent Claim
11 words
A recombinant host cell transformed with the vector of claim I2.
Claim 26
Dependent Claim
23 words
A method of producing DT-A-B' fragment which method comprises (a) culturing the recombinant cells of claim 23, and (b) recovering the DT-A-D' fragment.
References Cited (Backward Citations)
US4237224A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-05-15 | 1992-11-14 | 1993-05-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-05-15 | 1996-11-14 | 1997-05-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-05-15 | 2000-11-14 | 2001-05-16 | $7700.00 | Window Closed / Lapsed |