Patent US2983562A successfully refreshed from Google Patents.
US5047342A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Cloning and expression of T5 DNA polymerase
Assignee: Life Technologies Corp
•
Filing: 1989-08-10
•
Granted: 1991-09-10
•
Expires: 2009-08-10
Abstract
Abstract The present invention discloses a recombinant DNA molecule having a structural gene encoding a processive, thioredoxin-independent DNA polymerase, a promoter, and an origin of replication. The protein may also have a processive 3'-to-5' DNA exonuclease activity. A method for producing this enzyme is also disclosed, as is the protein produced by this process. This invention is exemplified by expression of T5 DNA polymerase in E. coli.
Inventors
- 👤 Deb K. Chatterjee
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 18:20
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/10; C12N9/12; C12N9/1241; C12N9/1252; Y; Y10; Y10S; Y10S530/00; Y10S530/82; Y10S530/826
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
76 words
An isolated recombinant DNA molecule comprising (a) a structural gene encoding a protein, wherein said protein has a T5 DNA polymerase activity, (b) a promoter, wherein said promoter and said structural gene are in such position and orientation with respect to each other that said structural gene may be expressed in a cell under control of said promoter, and (c) an origin of replication capable of maintaining the promoter/structural gene/origin of replication combination in a cell.
Claim 2
Dependent Claim
76 words
An isolated recombinant DNA molecule comprising (a) a structural gene encoding a protein, wherein said protein has a T5 DNA polymerase activity, (b) a promoter, wherein said promoter and said structural gene are in such position and orientation with respect to each other that said structural gene may be expressed in a cell under control of said promoter, and (c) an origin of replication capable of maintaining the promoter/structural gene/origin of replication combination in a cell.
Claim 3
Dependent Claim
17 words
A cell containing the molecule as in claim 1, wherein the molecule is contained by a cell.
Claim 4
Dependent Claim
14 words
The cell as in claim 2, wherein the cell is an E. coli cell.
Claim 5
Dependent Claim
11 words
The molecule as in claim 1, wherein the promoter is inducible.
Claim 6
Dependent Claim
16 words
The molecule as in claim 4, wherein the inducible promoter is a lambda p L promoter.
Claim 7
Dependent Claim
14 words
The molecule as in claim 4, wherein the inducible promoter is a tac promoter.
Claim 8
Dependent Claim
20 words
The molecule as in claim 1, wherein the structural gene encodes a protein having a processive 3'-to-5' DNA exonuclease activity.
Claim 9
Dependent Claim
17 words
The molecule as in claim 1, wherein the structural gene is under control of a heterologous promoter.
Claim 10
Dependent Claim
18 words
The molecule as in claim 1, wherein the structural gene is under control of a heterologous ribosome-binding site..
Claim 11
Dependent Claim
17 words
The molecule as in claim 1, wherein the origin of replication is heterologous to the structural gene.
Claim 12
Dependent Claim
20 words
The molecule as in claim 1, wherein the structural gene encodes a functional derivative of a T5 DNA polymerase protein.
Claim 13
Dependent Claim
21 words
The molecule as in claim 11, wherein the functional derivative has at least 90% homology with a T5 DNA polymerase protein.
Claim 14
Dependent Claim
21 words
The molecule as in claim 11, wherein the functional derivative has at least 75% homology with a T5 DNA polymerase protein.
Claim 15
Dependent Claim
11 words
The molecule as in claim 1, wherein the molecule is pTTQ19-T5-3.
Claim 16
Dependent Claim
15 words
The molecule as in claim 1, wherein the molecule is pTTQ19T-5-2 or a derivative thereof.
Claim 17
Dependent Claim
17 words
A cell containing the molecule of claim 15, wherein the cell is an E. coli BH215 cell.
Claim 18
Dependent Claim
28 words
The molecule as in claim 1, wherein the structural gene encodes a protein having T5 DNA polymerase activity synthesizes an average of at least 100 nucleotides before dissociation.
Claim 19
Dependent Claim
22 words
The molecule as in claim 17, wherein the T5 DNA polymerase activity synthesizes an average of at least 200 nucleotides before dissociation.
Claim 20
Dependent Claim
15 words
A recombinant DNA molecule, wherein the molecule is clone #2 as shown in FIG. 2.
Claim 21
Dependent Claim
15 words
A recombinant DNA molecule, wherein the molecule is clone #2 as shown in FIG. 2.
Claim 22
Dependent Claim
9 words
A recombinant DNA molecule, wherein the molecule is pTTQ19-T5-1.
Claim 23
Dependent Claim
9 words
A recombinant DNA molecule, wherein the molecule is pTTQ19-T5-1.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-09-09 | 1995-03-11 | 1995-09-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-09-09 | 1999-03-11 | 1999-09-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-09-09 | 2003-03-11 | 2003-09-10 | $7700.00 | Window Closed / Lapsed |