RU2197524C2 - Термостабильная днк-полимераза, способ ее получения, фрагмент выделенной днк, способ амплификации днк, способ введения метки в днк и способ обратной транскрипции - Google Patents
Термостабильная днк-полимераза, способ ее получения, фрагмент выделенной днк, способ амплификации днк, способ введения метки в днк и способ обратной транскрипции Download PDFInfo
- Publication number
- RU2197524C2 RU2197524C2 RU99109121/13A RU99109121A RU2197524C2 RU 2197524 C2 RU2197524 C2 RU 2197524C2 RU 99109121/13 A RU99109121/13 A RU 99109121/13A RU 99109121 A RU99109121 A RU 99109121A RU 2197524 C2 RU2197524 C2 RU 2197524C2
- Authority
- RU
- Russia
- Prior art keywords
- dna
- polymerase
- activity
- dna polymerase
- reverse transcriptase
- Prior art date
Links
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 title claims abstract description 97
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 title claims abstract description 96
- 238000000034 method Methods 0.000 title claims abstract description 48
- 239000012634 fragment Substances 0.000 title claims abstract description 20
- 238000010839 reverse transcription Methods 0.000 title claims abstract description 11
- 238000010348 incorporation Methods 0.000 title abstract description 3
- 230000004544 DNA amplification Effects 0.000 title description 2
- 230000000694 effects Effects 0.000 claims abstract description 81
- 241000620137 Carboxydothermus hydrogenoformans Species 0.000 claims abstract description 43
- 102100034343 Integrase Human genes 0.000 claims abstract description 36
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 claims abstract description 36
- 239000000872 buffer Substances 0.000 claims abstract description 21
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 claims abstract description 17
- 229910001425 magnesium ion Inorganic materials 0.000 claims abstract description 17
- 230000003321 amplification Effects 0.000 claims abstract description 10
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 10
- 229910001437 manganese ion Inorganic materials 0.000 claims abstract description 7
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 claims abstract description 6
- 230000001419 dependent effect Effects 0.000 claims abstract description 6
- 238000004587 chromatography analysis Methods 0.000 claims abstract 2
- 108020004414 DNA Proteins 0.000 claims description 75
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 9
- 239000011777 magnesium Substances 0.000 claims description 6
- 239000011572 manganese Substances 0.000 claims description 5
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims description 4
- 229910052749 magnesium Inorganic materials 0.000 claims description 4
- 238000000746 purification Methods 0.000 claims description 3
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims description 2
- 229920002684 Sepharose Polymers 0.000 claims description 2
- 229910052748 manganese Inorganic materials 0.000 claims description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims 1
- 238000006116 polymerization reaction Methods 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 abstract description 20
- 108090000790 Enzymes Proteins 0.000 abstract description 20
- 108060002716 Exonuclease Proteins 0.000 abstract description 18
- 102000013165 exonuclease Human genes 0.000 abstract description 18
- 239000013612 plasmid Substances 0.000 abstract description 15
- 239000002773 nucleotide Substances 0.000 abstract description 8
- 125000003729 nucleotide group Chemical group 0.000 abstract description 8
- 241000186394 Eubacterium Species 0.000 abstract 1
- 230000001747 exhibiting effect Effects 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 102000053602 DNA Human genes 0.000 description 71
- 239000013615 primer Substances 0.000 description 31
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 26
- 229920002477 rna polymer Polymers 0.000 description 24
- 108090000623 proteins and genes Proteins 0.000 description 22
- 150000007523 nucleic acids Chemical class 0.000 description 15
- 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 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 14
- 238000006243 chemical reaction Methods 0.000 description 13
- 239000012528 membrane Substances 0.000 description 13
- 239000011780 sodium chloride Substances 0.000 description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- 239000000499 gel Substances 0.000 description 12
- 230000014509 gene expression Effects 0.000 description 12
- 102000039446 nucleic acids Human genes 0.000 description 11
- 108020004707 nucleic acids Proteins 0.000 description 11
- 239000011159 matrix material Substances 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 239000013598 vector Substances 0.000 description 10
- 241000588724 Escherichia coli Species 0.000 description 9
- 239000002299 complementary DNA Substances 0.000 description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 8
- 238000011534 incubation Methods 0.000 description 8
- 210000004379 membrane Anatomy 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 239000001226 triphosphate Substances 0.000 description 8
- 235000011178 triphosphate Nutrition 0.000 description 8
- 241000620141 Carboxydothermus Species 0.000 description 7
- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 description 7
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 description 7
- SHIBSTMRCDJXLN-KCZCNTNESA-N digoxigenin Chemical compound C1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)C[C@H]3O)C)=CC(=O)OC1 SHIBSTMRCDJXLN-KCZCNTNESA-N 0.000 description 7
- 238000002955 isolation Methods 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 230000006820 DNA synthesis Effects 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 230000015556 catabolic process Effects 0.000 description 6
- 238000010367 cloning Methods 0.000 description 6
- 238000006731 degradation reaction Methods 0.000 description 6
- 238000001962 electrophoresis Methods 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 125000002264 triphosphate group Chemical class [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 6
- 239000004677 Nylon Substances 0.000 description 5
- 108091034117 Oligonucleotide Proteins 0.000 description 5
- 102100040853 PRKC apoptosis WT1 regulator protein Human genes 0.000 description 5
- 101710162991 PRKC apoptosis WT1 regulator protein Proteins 0.000 description 5
- 108020004511 Recombinant DNA Proteins 0.000 description 5
- 238000011065 in-situ storage Methods 0.000 description 5
- 229920001778 nylon Polymers 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 4
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 4
- 108010006785 Taq Polymerase Proteins 0.000 description 4
- 241000589500 Thermus aquaticus Species 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 244000309466 calf Species 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 125000004122 cyclic group Chemical group 0.000 description 4
- 239000005549 deoxyribonucleoside Substances 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 4
- 239000011976 maleic acid Substances 0.000 description 4
- -1 nucleoside triphosphates Chemical class 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 210000001541 thymus gland Anatomy 0.000 description 4
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 4
- 238000013518 transcription Methods 0.000 description 4
- 230000035897 transcription Effects 0.000 description 4
- 239000011534 wash buffer Substances 0.000 description 4
- AHCYMLUZIRLXAA-SHYZEUOFSA-N Deoxyuridine 5'-triphosphate Chemical compound O1[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C[C@@H]1N1C(=O)NC(=O)C=C1 AHCYMLUZIRLXAA-SHYZEUOFSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 241000205160 Pyrococcus Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 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
- 229960000723 ampicillin Drugs 0.000 description 3
- 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 3
- 239000000284 extract Substances 0.000 description 3
- 229930027917 kanamycin Natural products 0.000 description 3
- 229960000318 kanamycin Drugs 0.000 description 3
- 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 3
- 229930182823 kanamycin A Natural products 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 238000010369 molecular cloning Methods 0.000 description 3
- 229920002401 polyacrylamide Polymers 0.000 description 3
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 229920000136 polysorbate Polymers 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 238000004904 shortening Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 108020004635 Complementary DNA Proteins 0.000 description 2
- 102000012410 DNA Ligases Human genes 0.000 description 2
- 108010061982 DNA Ligases Proteins 0.000 description 2
- 230000004543 DNA replication Effects 0.000 description 2
- 238000001712 DNA sequencing Methods 0.000 description 2
- 102100031780 Endonuclease Human genes 0.000 description 2
- 108010042407 Endonucleases Proteins 0.000 description 2
- 108010067770 Endopeptidase K Proteins 0.000 description 2
- 101000638058 Homo sapiens Neurogenic differentiation factor 4 Proteins 0.000 description 2
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 2
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 2
- 102100032061 Neurogenic differentiation factor 4 Human genes 0.000 description 2
- 101710163270 Nuclease Proteins 0.000 description 2
- 102000006382 Ribonucleases Human genes 0.000 description 2
- 108010083644 Ribonucleases Proteins 0.000 description 2
- 102100035254 Sodium- and chloride-dependent GABA transporter 3 Human genes 0.000 description 2
- 101710104417 Sodium- and chloride-dependent GABA transporter 3 Proteins 0.000 description 2
- 241000589596 Thermus Species 0.000 description 2
- 241000589498 Thermus filiformis Species 0.000 description 2
- 241000589499 Thermus thermophilus Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 239000011543 agarose gel Substances 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000010804 cDNA synthesis Methods 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 239000013611 chromosomal DNA Substances 0.000 description 2
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 2
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 2
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 2
- UKWLRLAKGMZXJC-QIECWBMSSA-L disodium;[4-chloro-3-[(3r,5s)-1-chloro-3'-methoxyspiro[adamantane-4,4'-dioxetane]-3'-yl]phenyl] phosphate Chemical compound [Na+].[Na+].O1OC2([C@@H]3CC4C[C@H]2CC(Cl)(C4)C3)C1(OC)C1=CC(OP([O-])([O-])=O)=CC=C1Cl UKWLRLAKGMZXJC-QIECWBMSSA-L 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000012215 gene cloning Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 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 2
- 239000007788 liquid Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000002777 nucleoside Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000002028 premature Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 229940035044 sorbitan monolaurate Drugs 0.000 description 2
- 229920001187 thermosetting polymer Polymers 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- VLEIUWBSEKKKFX-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid Chemical compound OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O VLEIUWBSEKKKFX-UHFFFAOYSA-N 0.000 description 1
- WRDABNWSWOHGMS-UHFFFAOYSA-N AEBSF hydrochloride Chemical compound Cl.NCCC1=CC=C(S(F)(=O)=O)C=C1 WRDABNWSWOHGMS-UHFFFAOYSA-N 0.000 description 1
- 241000259810 Acremonium thermophilum Species 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 101000909256 Caldicellulosiruptor bescii (strain ATCC BAA-1888 / DSM 6725 / Z-1320) DNA polymerase I Proteins 0.000 description 1
- 229920002271 DEAE-Sepharose Polymers 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 102000007260 Deoxyribonuclease I Human genes 0.000 description 1
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 101710203526 Integrase Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 239000007993 MOPS buffer Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000003832 Nucleotidyltransferases Human genes 0.000 description 1
- 108090000119 Nucleotidyltransferases Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 1
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 101000902592 Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1) DNA polymerase Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 101710136739 Teichoic acid poly(glycerol phosphate) polymerase Proteins 0.000 description 1
- 108700009124 Transcription Initiation Site Proteins 0.000 description 1
- 108030003004 Triphosphatases Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 108010020713 Tth polymerase Proteins 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- QWXOJIDBSHLIFI-UHFFFAOYSA-N [3-(1-chloro-3'-methoxyspiro[adamantane-4,4'-dioxetane]-3'-yl)phenyl] dihydrogen phosphate Chemical compound O1OC2(C3CC4CC2CC(Cl)(C4)C3)C1(OC)C1=CC=CC(OP(O)(O)=O)=C1 QWXOJIDBSHLIFI-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000004252 chorionic villi Anatomy 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000012411 cloning technique Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 1
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 229940079920 digestives acid preparations Drugs 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000010249 in-situ analysis Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000001617 migratory effect Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000036438 mutation frequency Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 238000003906 pulsed field gel electrophoresis Methods 0.000 description 1
- 239000012521 purified sample Substances 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
- NLIVDORGVGAOOJ-MAHBNPEESA-M xylene cyanol Chemical compound [Na+].C1=C(C)C(NCC)=CC=C1C(\C=1C(=CC(OS([O-])=O)=CC=1)OS([O-])=O)=C\1C=C(C)\C(=[NH+]/CC)\C=C/1 NLIVDORGVGAOOJ-MAHBNPEESA-M 0.000 description 1
Images
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)
- C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
- Saccharide Compounds (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP96115873.0 | 1996-10-03 | ||
| EP96115873A EP0834569A1 (en) | 1996-10-03 | 1996-10-03 | Thermostable DNA polymerase from carboxydothermus hydrogenoformans |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| RU99109121A RU99109121A (ru) | 2001-05-20 |
| RU2197524C2 true RU2197524C2 (ru) | 2003-01-27 |
Family
ID=8223262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU99109121/13A RU2197524C2 (ru) | 1996-10-03 | 1997-10-01 | Термостабильная днк-полимераза, способ ее получения, фрагмент выделенной днк, способ амплификации днк, способ введения метки в днк и способ обратной транскрипции |
Country Status (12)
| Country | Link |
|---|---|
| US (1) | US6468775B1 (enExample) |
| EP (2) | EP0834569A1 (enExample) |
| JP (1) | JP4005637B2 (enExample) |
| AT (1) | ATE357527T1 (enExample) |
| AU (1) | AU717009B2 (enExample) |
| CA (1) | CA2268014C (enExample) |
| DE (1) | DE69737502T2 (enExample) |
| IL (1) | IL129085A (enExample) |
| NO (1) | NO323846B1 (enExample) |
| NZ (1) | NZ334652A (enExample) |
| RU (1) | RU2197524C2 (enExample) |
| WO (1) | WO1998014589A1 (enExample) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111733145A (zh) * | 2020-07-01 | 2020-10-02 | 济南国科医工科技发展有限公司 | 重组酶的纯化方法 |
| RU2809366C1 (ru) * | 2022-12-28 | 2023-12-11 | Федеральное бюджетное учреждение науки "Центральный научно-исследовательский институт эпидемиологии" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека (ФБУН ЦНИИ Эпидемиологии Роспотребнадзора) | Способ получения большого фрагмента Bst-полимеразы (варианты) |
Families Citing this family (27)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0921196A1 (en) * | 1997-12-02 | 1999-06-09 | Roche Diagnostics GmbH | Modified DNA-polymerase from carboxydothermus hydrogenoformans and its use for coupled reverse transcription and polymerase chain reaction |
| EP0922765B1 (en) * | 1997-12-02 | 2005-11-23 | Roche Diagnostics GmbH | Modified DNA-polymerase from carboxydothermus hydrogenoformans and its use for coupled reverse transcription and polymerase chain reaction |
| US20030228616A1 (en) * | 1999-10-29 | 2003-12-11 | Stratagene | DNA polymerase mutants with reverse transcriptase activity |
| US20040009486A1 (en) * | 1999-10-29 | 2004-01-15 | Sorge Joseph A. | Compositions and methods utilizing DNA polymerases |
| US20050123940A1 (en) * | 1999-10-29 | 2005-06-09 | Stratagene California | Compositions and methods for synthesizing cDNA |
| WO2006033665A1 (en) | 2004-03-16 | 2006-03-30 | Inist Inc. | Tat-based vaccine compositions and methods of making and using same |
| WO2003042383A1 (en) * | 2001-11-14 | 2003-05-22 | Toyo Boseki Kabushiki Kaisha | Dna synthesis promoters, dna polymerase-associated factors and utilization thereof |
| CA2520538C (en) | 2003-03-31 | 2014-04-29 | F. Hoffmann-La Roche Ag | Compositions and methods for detecting certain flaviviruses, including members of the japanese encephalitis virus serogroup |
| US8962293B2 (en) | 2006-10-18 | 2015-02-24 | Roche Molecular Systems, Inc. | DNA polymerases and related methods |
| WO2010111292A1 (en) | 2009-03-23 | 2010-09-30 | Nanirx, Inc. | Treatment of cancer with immunostimulatory hiv tat derivative polypeptides |
| JP5926247B2 (ja) | 2010-06-18 | 2016-05-25 | エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ | 増大した３’末端ミスマッチ識別能を有するｄｎａポリメラーゼ |
| EP2582803B1 (en) | 2010-06-18 | 2015-02-25 | Roche Diagnostics GmbH | Dna polymerases with increased 3'-mismatch discrimination |
| CA2802302C (en) | 2010-06-18 | 2016-04-26 | F. Hoffman-La Roche Ag | Dna polymerases with increased 3'-mismatch discrimination |
| JP5876477B2 (ja) | 2010-06-18 | 2016-03-02 | エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ | 増大した３’末端ミスマッチ識別能を有するｄｎａポリメラーゼ |
| EP2582802B1 (en) | 2010-06-18 | 2016-01-13 | Roche Diagnostics GmbH | Dna polymerases with increased 3'-mismatch discrimination |
| WO2011157438A1 (en) | 2010-06-18 | 2011-12-22 | Roche Diagnostics Gmbh | Dna polymerases with increasesed 3'-mismatch discrimination |
| EP2582804B1 (en) | 2010-06-18 | 2015-02-25 | Roche Diagnostics GmbH | Dna polymerases with increased 3'-mismatch discrimination |
| EP2582808B1 (en) | 2010-06-18 | 2015-04-22 | Roche Diagniostics GmbH | Dna polymerases with increased 3'-mismatch discrimination |
| EP2675897B1 (en) | 2011-02-15 | 2016-05-11 | Roche Diagnostics GmbH | Dna polymerases with increased 3'-mismatch discrimination |
| ES2561885T3 (es) | 2011-04-11 | 2016-03-01 | F. Hoffmann-La Roche Ag | ADN polimerasas de actividad mejorada |
| US9090883B2 (en) | 2011-07-28 | 2015-07-28 | Roche Molecular Systems, Inc. | DNA polymerases with improved activity |
| CN103987844B (zh) | 2011-12-08 | 2016-01-20 | 霍夫曼-拉罗奇有限公司 | 具有改进活性的dna聚合酶 |
| WO2013083262A1 (en) | 2011-12-08 | 2013-06-13 | Roche Diagnostics Gmbh | Dna polymerases with improved activity |
| WO2013083264A1 (en) | 2011-12-08 | 2013-06-13 | Roche Diagnostics Gmbh | Dna polymerases with improved activity |
| WO2015051245A1 (en) | 2013-10-04 | 2015-04-09 | Pin Pharma, Inc. | Immunostimulatory hiv tat derivative polypeptides for use in cancer treatment |
| WO2019180137A1 (en) | 2018-03-21 | 2019-09-26 | F. Hoffmann-La Roche Ag | Dna polymerases for efficient and effective incorporation of methylated-dntps |
| CN112703248B (zh) | 2018-09-13 | 2025-03-25 | 豪夫迈·罗氏有限公司 | 具有改良链置换能力的突变型dna聚合酶 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
| US5374553A (en) * | 1986-08-22 | 1994-12-20 | Hoffmann-La Roche Inc. | DNA encoding a thermostable nucleic acid polymerase enzyme from thermotoga maritima |
| US5912155A (en) * | 1994-09-30 | 1999-06-15 | Life Technologies, Inc. | Cloned DNA polymerases from Thermotoga neapolitana |
-
1996
- 1996-10-03 EP EP96115873A patent/EP0834569A1/en not_active Withdrawn
-
1997
- 1997-10-01 JP JP51624298A patent/JP4005637B2/ja not_active Expired - Fee Related
- 1997-10-01 IL IL129085A patent/IL129085A/en not_active IP Right Cessation
- 1997-10-01 AU AU50495/98A patent/AU717009B2/en not_active Ceased
- 1997-10-01 AT AT97913133T patent/ATE357527T1/de active
- 1997-10-01 RU RU99109121/13A patent/RU2197524C2/ru not_active IP Right Cessation
- 1997-10-01 NZ NZ334652A patent/NZ334652A/xx not_active IP Right Cessation
- 1997-10-01 CA CA002268014A patent/CA2268014C/en not_active Expired - Fee Related
- 1997-10-01 WO PCT/EP1997/005391 patent/WO1998014589A1/en not_active Ceased
- 1997-10-01 DE DE69737502T patent/DE69737502T2/de not_active Expired - Lifetime
- 1997-10-01 US US09/269,861 patent/US6468775B1/en not_active Expired - Lifetime
- 1997-10-01 EP EP97913133A patent/EP0929680B1/en not_active Expired - Lifetime
-
1999
- 1999-03-30 NO NO19991568A patent/NO323846B1/no not_active IP Right Cessation
Non-Patent Citations (1)
| Title |
|---|
| PERLER F.B et al. Thermostable DNA polymerases Advances in protein Chemistry, 1996, v.48, p.377-435. * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111733145A (zh) * | 2020-07-01 | 2020-10-02 | 济南国科医工科技发展有限公司 | 重组酶的纯化方法 |
| RU2809366C1 (ru) * | 2022-12-28 | 2023-12-11 | Федеральное бюджетное учреждение науки "Центральный научно-исследовательский институт эпидемиологии" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека (ФБУН ЦНИИ Эпидемиологии Роспотребнадзора) | Способ получения большого фрагмента Bst-полимеразы (варианты) |
| RU2815455C1 (ru) * | 2023-05-10 | 2024-03-18 | Федеральное государственное бюджетное учреждение науки Институт химической биологии и фундаментальной медицины Сибирского отделения Российской академии наук (ИХБФМ СО РАН) | Рекомбинантный штамм бактерий Escherichia coli Rosetta 2(DE3)/pET28c-Mau - продуцент ДНК-полимеразы А |
| RU2840236C1 (ru) * | 2024-07-26 | 2025-05-19 | Федеральное государственное бюджетное учреждение науки Институт химической биологии и фундаментальной медицины Сибирского отделения Российской академии наук (ИХБФМ СО РАН) | Рекомбинантный штамм бактерий Escherichia coli Rosetta 2(DE3)/pET28c-TstP36H-Sso7d - продуцент химерной термостабильной ДНК-полимеразы TstP36H-Sso7d из Thermococcus stetteri |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2001502170A (ja) | 2001-02-20 |
| NO323846B1 (no) | 2007-07-09 |
| CA2268014A1 (en) | 1998-04-09 |
| WO1998014589A1 (en) | 1998-04-09 |
| ATE357527T1 (de) | 2007-04-15 |
| DE69737502D1 (en) | 2007-05-03 |
| AU717009B2 (en) | 2000-03-16 |
| DE69737502T2 (de) | 2007-10-31 |
| NO991568L (no) | 1999-06-02 |
| AU5049598A (en) | 1998-04-24 |
| EP0929680B1 (en) | 2007-03-21 |
| NZ334652A (en) | 2000-03-27 |
| EP0929680A1 (en) | 1999-07-21 |
| NO991568D0 (no) | 1999-03-30 |
| US6468775B1 (en) | 2002-10-22 |
| EP0834569A1 (en) | 1998-04-08 |
| IL129085A (en) | 2006-12-10 |
| IL129085A0 (en) | 2000-02-17 |
| JP4005637B2 (ja) | 2007-11-07 |
| CA2268014C (en) | 2008-01-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| RU2197524C2 (ru) | Термостабильная днк-полимераза, способ ее получения, фрагмент выделенной днк, способ амплификации днк, способ введения метки в днк и способ обратной транскрипции | |
| JP4176890B2 (ja) | カルボキシドテルムス・ヒドロゲノフォルマンス由来の修飾ｄｎａポリメラーゼ並びに連結させた逆転写−ポリメラーゼ連鎖反応におけるその使用 | |
| WO1998014589A9 (en) | Thermostable dna polymerase from carboxydothermus hydrogenoformans | |
| JP3227101B2 (ja) | 熱安定性ｄｎａポリメラーゼ | |
| US5830714A (en) | Biologically active fragment of bacillus stearothermophilus DNA polymerase | |
| JPH07108220B2 (ja) | サーモトガ・マリチマ由来の熱安定性核酸ポリメラーゼ | |
| EP0799319B1 (en) | Purified thermococcus barossii dna polymerase | |
| RU2297454C2 (ru) | СПОСОБ АМПЛИФИКАЦИИ ДНК, СПОСОБ КЛОНИРОВАНИЯ ВТОРОЙ кДНК И СПОСОБ ОБРАТНОЙ ТРАНСКРИПЦИИ ДНК | |
| JP3132624B2 (ja) | 超好熱始原菌由来のｄｎａポリメラーゼ遺伝子およびその用途 | |
| EP0922765B1 (en) | Modified DNA-polymerase from carboxydothermus hydrogenoformans and its use for coupled reverse transcription and polymerase chain reaction | |
| WO2007076461A1 (en) | Thermostable dna polymerase from thermus scotoductus | |
| CA2252968C (en) | Modified dna-polymerase from carboxydothermus hydrogeno-formans and its use for coupled reverse transcription and polymerase chain reaction | |
| WO2007076464A2 (en) | Thermostable dna polymerase from thermus filiformis | |
| KR100372777B1 (ko) | 아퀴펙스 파이로필러스로부터 분리된 내열성 디엔에이중합효소의 유전자 및 그로부터 유추되는 아미노산 서열 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MM4A | The patent is invalid due to non-payment of fees |
Effective date: 20141002 |