CN114945663A - 耐高温逆转录酶突变体及其应用 - Google Patents
耐高温逆转录酶突变体及其应用 Download PDFInfo
- Publication number
- CN114945663A CN114945663A CN202080092826.9A CN202080092826A CN114945663A CN 114945663 A CN114945663 A CN 114945663A CN 202080092826 A CN202080092826 A CN 202080092826A CN 114945663 A CN114945663 A CN 114945663A
- Authority
- CN
- China
- Prior art keywords
- amino acid
- acid residue
- reverse transcriptase
- mutated
- 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.)
- Granted
Links
- 102100034343 Integrase Human genes 0.000 title claims abstract description 127
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 title claims abstract description 127
- 238000010839 reverse transcription Methods 0.000 claims abstract description 52
- 238000000034 method Methods 0.000 claims abstract description 36
- 125000000539 amino acid group Chemical group 0.000 claims description 180
- 230000035772 mutation Effects 0.000 claims description 52
- 210000004027 cell Anatomy 0.000 claims description 35
- 238000006243 chemical reaction Methods 0.000 claims description 27
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 19
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 15
- 241001529936 Murinae Species 0.000 claims description 14
- 208000032839 leukemia Diseases 0.000 claims description 13
- 239000013598 vector Substances 0.000 claims description 12
- 238000001514 detection method Methods 0.000 claims description 11
- 108091033319 polynucleotide Proteins 0.000 claims description 6
- 102000040430 polynucleotide Human genes 0.000 claims description 6
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 239000002157 polynucleotide Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 210000000349 chromosome Anatomy 0.000 claims description 2
- 238000012216 screening Methods 0.000 abstract description 26
- 230000003321 amplification Effects 0.000 abstract description 7
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 7
- 108090000623 proteins and genes Proteins 0.000 description 42
- 108090000790 Enzymes Proteins 0.000 description 40
- 102000004190 Enzymes Human genes 0.000 description 39
- 102000004169 proteins and genes Human genes 0.000 description 29
- 241000713869 Moloney murine leukemia virus Species 0.000 description 23
- 239000013612 plasmid Substances 0.000 description 13
- 230000000694 effects Effects 0.000 description 12
- 108020004414 DNA Proteins 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 108091028043 Nucleic acid sequence Proteins 0.000 description 9
- 238000003753 real-time PCR Methods 0.000 description 9
- 238000004587 chromatography analysis Methods 0.000 description 7
- 239000013604 expression vector Substances 0.000 description 7
- 230000006698 induction Effects 0.000 description 7
- 150000007523 nucleic acids Chemical class 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000002741 site-directed mutagenesis Methods 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000010804 cDNA synthesis Methods 0.000 description 5
- 238000007796 conventional method Methods 0.000 description 5
- 230000002209 hydrophobic effect Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 108020004707 nucleic acids Proteins 0.000 description 5
- 102000039446 nucleic acids Human genes 0.000 description 5
- 230000035484 reaction time Effects 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- 108020004635 Complementary DNA Proteins 0.000 description 4
- 102000053602 DNA Human genes 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- 108020004511 Recombinant DNA Proteins 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 239000012501 chromatography medium Substances 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 210000003527 eukaryotic cell Anatomy 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 238000001641 gel filtration chromatography Methods 0.000 description 4
- 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 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- 238000000108 ultra-filtration Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000001190 Q-PCR Methods 0.000 description 3
- 108020004682 Single-Stranded DNA Proteins 0.000 description 3
- 238000001042 affinity chromatography Methods 0.000 description 3
- 238000005571 anion exchange chromatography Methods 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229910021645 metal ion Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 210000001236 prokaryotic cell Anatomy 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 241000589158 Agrobacterium Species 0.000 description 2
- 241000713838 Avian myeloblastosis virus Species 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 241000711573 Coronaviridae Species 0.000 description 2
- 230000006820 DNA synthesis Effects 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 238000007846 asymmetric PCR Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000004520 electroporation Methods 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000005342 ion exchange Methods 0.000 description 2
- 239000012139 lysis buffer Substances 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000010188 recombinant method Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 210000005253 yeast cell Anatomy 0.000 description 2
- GUPXYSSGJWIURR-UHFFFAOYSA-N 3-octoxypropane-1,2-diol Chemical compound CCCCCCCCOCC(O)CO GUPXYSSGJWIURR-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 239000012619 Butyl Sepharose® Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000186226 Corynebacterium glutamicum Species 0.000 description 1
- 229920002271 DEAE-Sepharose Polymers 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 108060002716 Exonuclease Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 101710203526 Integrase Proteins 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 241000714177 Murine leukemia virus Species 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- 108010021466 Mutant Proteins Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000012614 Q-Sepharose Substances 0.000 description 1
- 108020004412 RNA 3' Polyadenylation Signals Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 239000012506 Sephacryl® Substances 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 239000012505 Superdex™ Substances 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 241000319304 [Brevibacterium] flavum Species 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000005277 cation exchange chromatography Methods 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000005189 flocculation Methods 0.000 description 1
- 230000016615 flocculation Effects 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229920002492 poly(sulfone) Polymers 0.000 description 1
- 239000013615 primer Substances 0.000 description 1
- 239000002987 primer (paints) Substances 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000013074 reference sample Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 230000011218 segmentation Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000002463 transducing effect Effects 0.000 description 1
- 238000003151 transfection method Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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)
- C12N9/1276—RNA-directed DNA polymerase (2.7.7.49), i.e. reverse transcriptase or telomerase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
-
- 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)
- C12Y207/07049—RNA-directed DNA polymerase (2.7.7.49), i.e. telomerase or reverse-transcriptase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/91—Transferases (2.)
- G01N2333/912—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- G01N2333/91205—Phosphotransferases in general
- G01N2333/91245—Nucleotidyltransferases (2.7.7)
- G01N2333/9125—Nucleotidyltransferases (2.7.7) with a definite EC number (2.7.7.-)
- G01N2333/9128—RNA-directed DNA polymerases, e.g. RT (2.7.7.49)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明提供了耐高温逆转录酶突变体及其应用，具体地，本发明构建了逆转录酶(M‑MLV)突变库，通过逐步筛选，最终筛选出了热稳定性提高且扩增效率较高的突变体。在高温条件下，与野生型相比，本发明的逆转录酶突变体的逆转录效率得到了极大的提高。
Description
PCT国内申请，说明书已公开。
Claims (13)
- PCT国内申请，权利要求书已公开。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911105358.XA CN112795551B (zh) | 2019-11-13 | 2019-11-13 | 一种耐高温逆转录酶突变体及其应用 |
| CN201911105358X | 2019-11-13 | ||
| PCT/CN2020/128639 WO2021093845A1 (zh) | 2019-11-13 | 2020-11-13 | 耐高温逆转录酶突变体及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114945663A true CN114945663A (zh) | 2022-08-26 |
| CN114945663B CN114945663B (zh) | 2024-05-31 |
Family
ID=
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001092500A1 (en) * | 2000-05-26 | 2001-12-06 | Invitrogen Corporation | Thermostable reverse transcriptases and uses thereof |
| US20020090618A1 (en) * | 2000-05-26 | 2002-07-11 | Invitrogen Corporation | Thermostable reverse transcriptases and uses thereof |
| CN1422336A (zh) * | 2000-03-15 | 2003-06-04 | 茵维特罗根公司 | 高保真逆转录酶及其用途 |
| WO2007022045A2 (en) * | 2005-08-10 | 2007-02-22 | Stratagene California | Mutant reverse transcriptase and methods of use |
| US20110065606A1 (en) * | 2008-04-10 | 2011-03-17 | Fermentas Uab | Production of nucleic acid |
| US20130143225A1 (en) * | 2010-08-13 | 2013-06-06 | Kyoto University | Variant reverse transcriptase |
| CN106164261A (zh) * | 2014-01-22 | 2016-11-23 | 生命技术公司 | 适用于高温核酸合成的新颖逆转录酶 |
| CN106906237A (zh) * | 2017-04-18 | 2017-06-30 | 淮海工学院 | 一种高性能m‑mlv逆转录酶的制备方法 |
| US20180312822A1 (en) * | 2017-04-26 | 2018-11-01 | 10X Genomics, Inc. | Mmlv reverse transcriptase variants |
| CN114480337A (zh) * | 2020-11-13 | 2022-05-13 | 广州达安基因股份有限公司 | 逆转录酶突变体及逆转录方法 |
| CN114480329A (zh) * | 2020-11-13 | 2022-05-13 | 广州达安基因股份有限公司 | 高效率mmlv酶突变体 |
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1422336A (zh) * | 2000-03-15 | 2003-06-04 | 茵维特罗根公司 | 高保真逆转录酶及其用途 |
| WO2001092500A1 (en) * | 2000-05-26 | 2001-12-06 | Invitrogen Corporation | Thermostable reverse transcriptases and uses thereof |
| US20020090618A1 (en) * | 2000-05-26 | 2002-07-11 | Invitrogen Corporation | Thermostable reverse transcriptases and uses thereof |
| CN1430670A (zh) * | 2000-05-26 | 2003-07-16 | 茵维特罗根公司 | 热稳定逆转录酶及其用途 |
| WO2007022045A2 (en) * | 2005-08-10 | 2007-02-22 | Stratagene California | Mutant reverse transcriptase and methods of use |
| CN102057039A (zh) * | 2008-04-10 | 2011-05-11 | 菲门特斯Uab公司 | 核酸的制备 |
| US20110065606A1 (en) * | 2008-04-10 | 2011-03-17 | Fermentas Uab | Production of nucleic acid |
| CN107058258A (zh) * | 2008-04-10 | 2017-08-18 | 赛默飞世尔科技波罗的海Uvb公司 | 一种逆转录酶和编码其的多核苷酸 |
| US20130143225A1 (en) * | 2010-08-13 | 2013-06-06 | Kyoto University | Variant reverse transcriptase |
| CN106164261A (zh) * | 2014-01-22 | 2016-11-23 | 生命技术公司 | 适用于高温核酸合成的新颖逆转录酶 |
| CN106906237A (zh) * | 2017-04-18 | 2017-06-30 | 淮海工学院 | 一种高性能m‑mlv逆转录酶的制备方法 |
| US20180312822A1 (en) * | 2017-04-26 | 2018-11-01 | 10X Genomics, Inc. | Mmlv reverse transcriptase variants |
| CN114480337A (zh) * | 2020-11-13 | 2022-05-13 | 广州达安基因股份有限公司 | 逆转录酶突变体及逆转录方法 |
| CN114480329A (zh) * | 2020-11-13 | 2022-05-13 | 广州达安基因股份有限公司 | 高效率mmlv酶突变体 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112795551A (zh) | 2021-05-14 |
| CN112795551B (zh) | 2024-01-30 |
| EP3848458A1 (en) | 2021-07-14 |
| EP3848458A4 (en) | 2022-11-30 |
| WO2021093845A1 (zh) | 2021-05-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112795551B (zh) | 一种耐高温逆转录酶突变体及其应用 | |
| CN111484987B (zh) | 一种具有高扩增活性的耐热dna聚合酶突变体 | |
| CN112795546B (zh) | 一种具有高逆转录效率的耐高温逆转录酶突变体及其应用 | |
| CN112795548A (zh) | 具有高逆转录效率的耐高温逆转录酶突变体及其应用 | |
| CN114480334B (zh) | 用于新冠病毒检测的逆转录酶突变体 | |
| CN112795549B (zh) | 一种逆转录酶突变体 | |
| CN115485374A (zh) | 一种具有高扩增活性的耐热dna聚合酶突变体 | |
| CN114945663B (zh) | 耐高温逆转录酶突变体及其应用 | |
| CN114480332B (zh) | 耐高温m-mlv酶突变体及其应用 | |
| CN114480336B (zh) | 含有逆转录酶突变体的核酸检测试剂盒 | |
| CN114480333B (zh) | 逆转录酶突变体及其应用 | |
| CN114480338B (zh) | 用于核酸检测的逆转录酶突变体 | |
| CN114480329B (zh) | 高效率mmlv酶突变体 | |
| CN114480337B (zh) | 逆转录酶突变体及逆转录方法 | |
| CN112795550B (zh) | 耐高温逆转录酶突变体 | |
| CN114480330A (zh) | 多突变位点的逆转录酶突变体 | |
| CN112795547B (zh) | 高逆转录效率的逆转录酶突变体 | |
| CN114480335A (zh) | 逆转录酶及逆转录检测试剂 | |
| CN114480331A (zh) | Mmlv酶组合突变体 | |
| CN112795547A (zh) | 高逆转录效率的逆转录酶突变体 | |
| CN112795550A (zh) | 耐高温逆转录酶突变体 | |
| CN112779238B (zh) | 用于丙肝病毒检测的dna聚合酶混合物 | |
| CN112592905B (zh) | 用于新型冠状病毒检测的dna聚合酶混合物 | |
| CN112779237B (zh) | 用于乙肝病毒检测的dna聚合酶混合物 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant |