ES2242733T3 - Mutante de arn polimerasa con termoestabilidad mejorada. - Google Patents
Mutante de arn polimerasa con termoestabilidad mejorada.Info
- Publication number
- ES2242733T3 ES2242733T3 ES01909810T ES01909810T ES2242733T3 ES 2242733 T3 ES2242733 T3 ES 2242733T3 ES 01909810 T ES01909810 T ES 01909810T ES 01909810 T ES01909810 T ES 01909810T ES 2242733 T3 ES2242733 T3 ES 2242733T3
- Authority
- ES
- Spain
- Prior art keywords
- rna polymerase
- baselineskip
- rna
- gene
- mutant
- 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
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 title claims description 47
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 title claims description 47
- 230000035772 mutation Effects 0.000 claims abstract description 46
- 101710137500 T7 RNA polymerase Proteins 0.000 claims abstract description 41
- 108090000623 proteins and genes Proteins 0.000 claims description 42
- 238000013518 transcription Methods 0.000 claims description 38
- 230000035897 transcription Effects 0.000 claims description 38
- 102000004190 Enzymes Human genes 0.000 claims description 36
- 108090000790 Enzymes Proteins 0.000 claims description 36
- 230000000694 effects Effects 0.000 claims description 32
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 25
- 230000003321 amplification Effects 0.000 claims description 24
- 238000006243 chemical reaction Methods 0.000 claims description 21
- 239000013598 vector Substances 0.000 claims description 20
- 150000007523 nucleic acids Chemical class 0.000 claims description 16
- 230000014509 gene expression Effects 0.000 claims description 14
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- 102000039446 nucleic acids Human genes 0.000 claims description 10
- 150000001413 amino acids Chemical class 0.000 claims description 9
- 102100034343 Integrase Human genes 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 6
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- 108010035563 Chloramphenicol O-acetyltransferase Proteins 0.000 description 4
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- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
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- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 4
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- 229920002873 Polyethylenimine Polymers 0.000 description 3
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 108010028263 bacteriophage T3 RNA polymerase Proteins 0.000 description 3
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- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
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- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
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- 241000193830 Bacillus <bacterium> Species 0.000 description 1
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- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 101100283604 Caenorhabditis elegans pigk-1 gene Proteins 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
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- 241000701867 Enterobacteria phage T7 Species 0.000 description 1
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- 108060002716 Exonuclease Proteins 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
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- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 description 1
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- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
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- 230000009948 RNA mutation Effects 0.000 description 1
- 108010065868 RNA polymerase SP6 Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
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- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 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 1
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- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 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 1
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- NJTGANWAUPEOAX-UHFFFAOYSA-N molport-023-220-454 Chemical compound OCC(O)CO.OCC(O)CO NJTGANWAUPEOAX-UHFFFAOYSA-N 0.000 description 1
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- QGFSVPWZEPKNDV-BRTFOEFASA-N ranp Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CSSC[C@@H](C(=O)N1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)=O)[C@@H](C)CC)[C@@H](C)CC)C1=CC=CC=C1 QGFSVPWZEPKNDV-BRTFOEFASA-N 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
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- 125000002652 ribonucleotide group Chemical group 0.000 description 1
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- 239000001226 triphosphate Substances 0.000 description 1
- 235000011178 triphosphate Nutrition 0.000 description 1
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1241—Nucleotidyltransferases (2.7.7)
- C12N9/1247—DNA-directed RNA polymerase (2.7.7.6)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
- C12Y207/07—Nucleotidyltransferases (2.7.7)
- C12Y207/07006—DNA-directed RNA polymerase (2.7.7.6)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Heat Sensitive Colour Forming Recording (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP00200787 | 2000-03-07 | ||
| EP00200787 | 2000-03-07 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2242733T3 true ES2242733T3 (es) | 2005-11-16 |
Family
ID=8171152
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES01909810T Expired - Lifetime ES2242733T3 (es) | 2000-03-07 | 2001-03-01 | Mutante de arn polimerasa con termoestabilidad mejorada. |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US7507567B2 (enExample) |
| EP (1) | EP1261696B1 (enExample) |
| JP (1) | JP4832694B2 (enExample) |
| AT (1) | ATE294855T1 (enExample) |
| AU (1) | AU2001237425A1 (enExample) |
| CA (1) | CA2400049C (enExample) |
| DE (1) | DE60110564T2 (enExample) |
| ES (1) | ES2242733T3 (enExample) |
| WO (1) | WO2001066705A1 (enExample) |
Families Citing this family (24)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009136153A (ja) * | 2007-12-03 | 2009-06-25 | Tosoh Corp | 変異型ｒｎａポリメラーゼ |
| WO2009113718A1 (ja) * | 2008-03-11 | 2009-09-17 | 東ソー株式会社 | 機能改善したｒｎａポリメラーゼ変異体 |
| KR20110044753A (ko) * | 2008-08-08 | 2011-04-29 | 토소가부시키가이샤 | 기능 개선된 ｒｎａ 중합효소 변이체 |
| US9193959B2 (en) * | 2010-04-16 | 2015-11-24 | Roche Diagnostics Operations, Inc. | T7 RNA polymerase variants with enhanced thermostability |
| DK3219796T3 (da) | 2010-08-31 | 2020-12-21 | Greenlight Biosciences Inc | Fremgangsmåder til kontrol af strømningen i metaboliske reaktionsveje ved hjælp af proteasemanipulation |
| US9062292B2 (en) | 2010-09-13 | 2015-06-23 | Enzo Life Sciences Inc. | Mutant T7 polymerases |
| FR2981088B1 (fr) | 2011-10-06 | 2013-11-29 | Biomerieux Sa | Arn polymerases mutees |
| US9045740B2 (en) * | 2012-02-24 | 2015-06-02 | University Of Massachusetts | Modified T7-related RNA polymerases and methods of use thereof |
| US9688977B2 (en) | 2013-08-05 | 2017-06-27 | Greenlight Biosciences, Inc. | Engineered phosphoglucose isomerase proteins with a protease cleavage site |
| WO2015024017A2 (en) * | 2013-08-16 | 2015-02-19 | President And Fellows Of Harvard College | Rna polymerase, methods of purification and methods of use |
| KR20160132113A (ko) * | 2014-03-20 | 2016-11-16 | 보드 오브 리전츠, 더 유니버시티 오브 텍사스 시스템 | 확장된 기질 범위 및 증진된 전사 수율을 갖는 t7 rna 폴리머라제 변이체 |
| AU2016243623B2 (en) | 2015-03-30 | 2022-04-14 | Greenlight Biosciences, Inc. | Cell-free production of ribonucleic acid |
| US20190017100A1 (en) * | 2015-07-01 | 2019-01-17 | Curevac Ag | Method for analysis of an rna molecule |
| CN108779446B (zh) | 2016-01-13 | 2023-04-21 | 新英格兰生物实验室公司 | T7 rna聚合酶的热稳定变体 |
| EP3440215A1 (en) | 2016-04-06 | 2019-02-13 | Greenlight Biosciences, Inc. | Cell-free production of ribonucleic acid |
| US10034951B1 (en) | 2017-06-21 | 2018-07-31 | New England Biolabs, Inc. | Use of thermostable RNA polymerases to produce RNAs having reduced immunogenicity |
| JP2020530267A (ja) | 2017-06-30 | 2020-10-22 | コデクシス， インコーポレイテッド | Ｔ７ ｒｎａポリメラーゼバリアント |
| EP3645711A4 (en) | 2017-06-30 | 2021-04-21 | Codexis, Inc. | T7 RNA POLYMERASE VARIANTS |
| AR113764A1 (es) | 2017-10-11 | 2020-06-10 | Greenlight Biosciences Inc | Métodos y composiciones para la producción de nucleósido trifosfato y ácido ribonucleico |
| AU2019253693A1 (en) * | 2018-04-10 | 2020-10-22 | Greenlight Biosciences, Inc. | T7 RNA polymerase variants |
| WO2020243026A1 (en) * | 2019-05-24 | 2020-12-03 | Primordial Genetics Inc. | Methods and compositions for manufacturing polynucleotides |
| CN112980811B (zh) * | 2021-03-06 | 2022-02-25 | 苏州瀚源新酶生物科技有限公司 | Rna聚合酶突变体及其应用、重组载体及其制备方法和应用、重组工程菌及其应用 |
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Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4952496A (en) * | 1984-03-30 | 1990-08-28 | Associated Universities, Inc. | Cloning and expression of the gene for bacteriophage T7 RNA polymerase |
| CA1340807C (en) * | 1988-02-24 | 1999-11-02 | Lawrence T. Malek | Nucleic acid amplification process |
| US5385834A (en) * | 1993-08-13 | 1995-01-31 | Georgia Tech Research Corporation | Mutant T7 RNA polymerase GP1(lys222) exhibiting altered promoter recognition |
| FR2761695B1 (fr) | 1997-04-04 | 1999-09-24 | Bio Merieux | Arn polymerase fonctionnant preferentiellement sur matrice d'arn, sa preparation et son utilisation |
| JP3172710B2 (ja) | 1997-07-07 | 2001-06-04 | 理化学研究所 | Ｒｎａポリメラーゼ |
| EP1137781B1 (en) | 1998-12-11 | 2004-05-12 | bioMerieux B.V. | Mutants of rna polymerases with increased stability |
| JP4399684B2 (ja) * | 1999-08-17 | 2010-01-20 | 東洋紡績株式会社 | 改良されたｒｎａポリメラーゼ |
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| JP4832694B2 (ja) | 2011-12-07 |
| CA2400049A1 (en) | 2001-09-13 |
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| EP1261696B1 (en) | 2005-05-04 |
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| JP2003525627A (ja) | 2003-09-02 |
| CA2400049C (en) | 2010-10-05 |
| DE60110564T2 (de) | 2006-01-19 |
| ATE294855T1 (de) | 2005-05-15 |
| DE60110564D1 (en) | 2005-06-09 |
| AU2001237425A1 (en) | 2001-09-17 |
| EP1261696A1 (en) | 2002-12-04 |
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