CA1335263C - Polymerase t7 de l'adn - Google Patents
Polymerase t7 de l'adnInfo
- Publication number
- CA1335263C CA1335263C CA000616696A CA616696A CA1335263C CA 1335263 C CA1335263 C CA 1335263C CA 000616696 A CA000616696 A CA 000616696A CA 616696 A CA616696 A CA 616696A CA 1335263 C CA1335263 C CA 1335263C
- Authority
- CA
- Canada
- Prior art keywords
- polymerase
- dna
- exonuclease activity
- fragment
- primer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 title claims abstract description 122
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 title claims abstract description 122
- 108020004414 DNA Proteins 0.000 claims abstract description 148
- 230000000694 effects Effects 0.000 claims abstract description 90
- 238000000034 method Methods 0.000 claims abstract description 73
- 239000012634 fragment Substances 0.000 claims abstract description 65
- 108060002716 Exonuclease Proteins 0.000 claims abstract description 58
- 102000013165 exonuclease Human genes 0.000 claims abstract description 58
- 102000053602 DNA Human genes 0.000 claims abstract description 40
- 238000000338 in vitro Methods 0.000 claims abstract description 11
- 238000002703 mutagenesis Methods 0.000 claims abstract description 11
- 231100000350 mutagenesis Toxicity 0.000 claims abstract description 11
- 238000004519 manufacturing process Methods 0.000 claims abstract description 9
- 125000003729 nucleotide group Chemical group 0.000 claims description 36
- 239000002773 nucleotide Substances 0.000 claims description 35
- 239000000203 mixture Substances 0.000 claims description 20
- 238000000137 annealing Methods 0.000 claims description 19
- 238000010348 incorporation Methods 0.000 claims description 11
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 10
- 230000015572 biosynthetic process Effects 0.000 claims description 10
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- 238000003199 nucleic acid amplification method Methods 0.000 claims description 8
- 108020004682 Single-Stranded DNA Proteins 0.000 claims description 7
- 239000005546 dideoxynucleotide Substances 0.000 claims description 6
- 238000011534 incubation Methods 0.000 claims description 5
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- 230000002194 synthesizing effect Effects 0.000 claims description 5
- 238000013467 fragmentation Methods 0.000 claims 2
- 238000006062 fragmentation reaction Methods 0.000 claims 2
- 241000701867 Enterobacteria phage T7 Species 0.000 claims 1
- 238000006243 chemical reaction Methods 0.000 description 58
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- 230000014509 gene expression Effects 0.000 description 9
- 125000002264 triphosphate group Chemical class [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 9
- 101150118060 trxA gene Proteins 0.000 description 9
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- UCBPDSYUVAAHCD-UWVGGRQHSA-N Leu-Pro-Gly Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UCBPDSYUVAAHCD-UWVGGRQHSA-N 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 8
- CCDFBRZVTDDJNM-GUBZILKMSA-N Ala-Leu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O CCDFBRZVTDDJNM-GUBZILKMSA-N 0.000 description 7
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 7
- -1 nucleotide triphosphates Chemical class 0.000 description 7
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- 238000007254 oxidation reaction Methods 0.000 description 7
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- 238000010561 standard procedure Methods 0.000 description 7
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 6
- DSFYPIUSAMSERP-IHRRRGAJSA-N Leu-Leu-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DSFYPIUSAMSERP-IHRRRGAJSA-N 0.000 description 6
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 6
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- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 6
- 229910052742 iron Inorganic materials 0.000 description 6
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- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 6
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 5
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- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 5
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- WALVCOOOKULCQM-ULQDDVLXSA-N Lys-Arg-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O WALVCOOOKULCQM-ULQDDVLXSA-N 0.000 description 3
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- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 3
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA000616696A CA1335263C (fr) | 1987-01-14 | 1993-08-12 | Polymerase t7 de l'adn |
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US07/003,227 US4795699A (en) | 1987-01-14 | 1987-01-14 | T7 DNA polymerase |
| US3,227 | 1987-01-14 | ||
| US132,569 | 1987-12-14 | ||
| US07/132,569 US4942130A (en) | 1987-01-14 | 1987-12-14 | T7 DNA polymerase |
| CA000556390A CA1340628C (fr) | 1987-01-14 | 1988-01-13 | Polymerase d'adn de type t7 |
| CA000616696A CA1335263C (fr) | 1987-01-14 | 1993-08-12 | Polymerase t7 de l'adn |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000616696A Division CA1335263C (fr) | 1987-01-14 | 1993-08-12 | Polymerase t7 de l'adn |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000616696A Division CA1335263C (fr) | 1987-01-14 | 1993-08-12 | Polymerase t7 de l'adn |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1335263C true CA1335263C (fr) | 1995-04-18 |
Family
ID=27167848
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000616696A Expired - Lifetime CA1335263C (fr) | 1987-01-14 | 1993-08-12 | Polymerase t7 de l'adn |
| CA000616697A Expired - Lifetime CA1335962C (fr) | 1987-01-14 | 1993-08-12 | Methode pour bloquer l'activite de l'exonuclease |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000616697A Expired - Lifetime CA1335962C (fr) | 1987-01-14 | 1993-08-12 | Methode pour bloquer l'activite de l'exonuclease |
Country Status (1)
| Country | Link |
|---|---|
| CA (2) | CA1335263C (fr) |
-
1993
- 1993-08-12 CA CA000616696A patent/CA1335263C/fr not_active Expired - Lifetime
- 1993-08-12 CA CA000616697A patent/CA1335962C/fr not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| CA1335962C (fr) | 1995-06-20 |
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Legal Events
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| MKEX | Expiry |