CA3024184A1 - Variants of a dna polymerase of the polx family - Google Patents
Variants of a dna polymerase of the polx family Download PDFInfo
- Publication number
- CA3024184A1 CA3024184A1 CA3024184A CA3024184A CA3024184A1 CA 3024184 A1 CA3024184 A1 CA 3024184A1 CA 3024184 A CA3024184 A CA 3024184A CA 3024184 A CA3024184 A CA 3024184A CA 3024184 A1 CA3024184 A1 CA 3024184A1
- Authority
- CA
- Canada
- Prior art keywords
- variant
- residue
- dna polymerase
- seq
- nucleic acid
- 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.)
- Pending
Links
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 title claims abstract description 91
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 title claims abstract description 91
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 100
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 74
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 70
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 70
- 101100191004 Bacillus subtilis (strain 168) polX gene Proteins 0.000 claims abstract description 54
- 230000035772 mutation Effects 0.000 claims abstract description 46
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 31
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 27
- 239000012634 fragment Substances 0.000 claims abstract description 15
- 239000002773 nucleotide Substances 0.000 claims description 58
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 claims description 46
- 238000006467 substitution reaction Methods 0.000 claims description 45
- 230000002194 synthesizing effect Effects 0.000 claims description 24
- 239000013598 vector Substances 0.000 claims description 24
- 125000000539 amino acid group Chemical group 0.000 claims description 15
- 238000012217 deletion Methods 0.000 claims description 15
- 230000037430 deletion Effects 0.000 claims description 15
- 230000002255 enzymatic effect Effects 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 14
- 108020004414 DNA Proteins 0.000 claims description 13
- 229910052727 yttrium Inorganic materials 0.000 claims description 11
- 229910052717 sulfur Inorganic materials 0.000 claims description 10
- 229910052739 hydrogen Inorganic materials 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 9
- 102000053602 DNA Human genes 0.000 claims description 7
- 229910052731 fluorine Inorganic materials 0.000 claims description 7
- 229910052698 phosphorus Inorganic materials 0.000 claims description 6
- 229910052700 potassium Inorganic materials 0.000 claims description 5
- 239000001587 sorbitan monostearate Substances 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 2
- 238000007792 addition Methods 0.000 description 57
- 210000004027 cell Anatomy 0.000 description 46
- 239000000872 buffer Substances 0.000 description 45
- 238000010348 incorporation Methods 0.000 description 34
- 102000004190 Enzymes Human genes 0.000 description 33
- 108090000790 Enzymes Proteins 0.000 description 33
- 150000001413 amino acids Chemical class 0.000 description 23
- 230000000694 effects Effects 0.000 description 21
- 238000005070 sampling Methods 0.000 description 21
- 238000005406 washing Methods 0.000 description 20
- 235000001014 amino acid Nutrition 0.000 description 19
- 229940024606 amino acid Drugs 0.000 description 19
- 238000010511 deprotection reaction Methods 0.000 description 19
- 239000000203 mixture Substances 0.000 description 19
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 16
- 238000012360 testing method Methods 0.000 description 14
- 230000003197 catalytic effect Effects 0.000 description 12
- 239000013612 plasmid Substances 0.000 description 10
- 239000001226 triphosphate Substances 0.000 description 10
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- 230000004048 modification Effects 0.000 description 8
- 238000012986 modification Methods 0.000 description 8
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 241000282414 Homo sapiens Species 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 5
- 241000206602 Eukaryota Species 0.000 description 5
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 5
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 5
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 5
- 239000011324 bead Substances 0.000 description 5
- 238000010828 elution Methods 0.000 description 5
- 210000004962 mammalian cell Anatomy 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 239000008188 pellet Substances 0.000 description 5
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 240000000594 Heliconia bihai Species 0.000 description 4
- 241000282412 Homo Species 0.000 description 4
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 4
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 4
- 241001529936 Murinae Species 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- 108010076504 Protein Sorting Signals Proteins 0.000 description 4
- 235000009582 asparagine Nutrition 0.000 description 4
- 229960001230 asparagine Drugs 0.000 description 4
- 235000003704 aspartic acid Nutrition 0.000 description 4
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 4
- 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 4
- 229960005091 chloramphenicol Drugs 0.000 description 4
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 4
- 230000001747 exhibiting effect Effects 0.000 description 4
- 229930027917 kanamycin Natural products 0.000 description 4
- 229960000318 kanamycin Drugs 0.000 description 4
- 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 4
- 229930182823 kanamycin A Natural products 0.000 description 4
- 239000012139 lysis buffer Substances 0.000 description 4
- 230000005012 migration Effects 0.000 description 4
- 238000013508 migration Methods 0.000 description 4
- 238000002703 mutagenesis Methods 0.000 description 4
- 231100000350 mutagenesis Toxicity 0.000 description 4
- 229920002401 polyacrylamide Polymers 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000002601 radiography Methods 0.000 description 4
- 239000011541 reaction mixture Substances 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000004475 Arginine Substances 0.000 description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 3
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 3
- 241000282577 Pan troglodytes Species 0.000 description 3
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 3
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 3
- 239000004473 Threonine Substances 0.000 description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 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 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 229930182817 methionine Natural products 0.000 description 3
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 3
- 108091033319 polynucleotide Proteins 0.000 description 3
- 102000040430 polynucleotide Human genes 0.000 description 3
- 239000002157 polynucleotide Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000001308 synthesis method Methods 0.000 description 3
- 230000008685 targeting Effects 0.000 description 3
- 239000013603 viral vector Substances 0.000 description 3
- 239000011534 wash buffer Substances 0.000 description 3
- -1 6-N-methyllysine Chemical compound 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 102100033215 DNA nucleotidylexotransferase Human genes 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 241001138401 Kluyveromyces lactis Species 0.000 description 2
- 241000235058 Komagataella pastoris Species 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
- 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 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- KSPIYJQBLVDRRI-UHFFFAOYSA-N N-methylisoleucine Chemical compound CCC(C)C(NC)C(O)=O KSPIYJQBLVDRRI-UHFFFAOYSA-N 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Chemical compound CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000002572 peristaltic effect Effects 0.000 description 2
- 230000002285 radioactive effect Effects 0.000 description 2
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical compound C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 description 2
- 239000011686 zinc sulphate Substances 0.000 description 2
- 235000009529 zinc sulphate Nutrition 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- BVAUMRCGVHUWOZ-ZETCQYMHSA-N (2s)-2-(cyclohexylazaniumyl)propanoate Chemical compound OC(=O)[C@H](C)NC1CCCCC1 BVAUMRCGVHUWOZ-ZETCQYMHSA-N 0.000 description 1
- OCUSNPIJIZCRSZ-ZTZWCFDHSA-N (2s)-2-amino-3-methylbutanoic acid;(2s)-2-amino-4-methylpentanoic acid;(2s,3s)-2-amino-3-methylpentanoic acid Chemical compound CC(C)[C@H](N)C(O)=O.CC[C@H](C)[C@H](N)C(O)=O.CC(C)C[C@H](N)C(O)=O OCUSNPIJIZCRSZ-ZTZWCFDHSA-N 0.000 description 1
- NPWMTBZSRRLQNJ-VKHMYHEASA-N (3s)-3-aminopiperidine-2,6-dione Chemical compound N[C@H]1CCC(=O)NC1=O NPWMTBZSRRLQNJ-VKHMYHEASA-N 0.000 description 1
- OSBLTNPMIGYQGY-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid;boric acid Chemical compound OB(O)O.OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O OSBLTNPMIGYQGY-UHFFFAOYSA-N 0.000 description 1
- 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 1
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 102000001421 BRCT domains Human genes 0.000 description 1
- 108050009608 BRCT domains Proteins 0.000 description 1
- 241000867607 Chlorocebus sabaeus Species 0.000 description 1
- 230000008265 DNA repair mechanism Effects 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000701832 Enterobacteria phage T3 Species 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108091092584 GDNA Proteins 0.000 description 1
- 101100424825 Gallus gallus DNTT gene Proteins 0.000 description 1
- 101000800646 Homo sapiens DNA nucleotidylexotransferase Proteins 0.000 description 1
- LCWXJXMHJVIJFK-UHFFFAOYSA-N Hydroxylysine Natural products NCC(O)CC(N)CC(O)=O LCWXJXMHJVIJFK-UHFFFAOYSA-N 0.000 description 1
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 1
- SNDPXSYFESPGGJ-BYPYZUCNSA-N L-2-aminopentanoic acid Chemical compound CCC[C@H](N)C(O)=O SNDPXSYFESPGGJ-BYPYZUCNSA-N 0.000 description 1
- AGPKZVBTJJNPAG-UHNVWZDZSA-N L-allo-Isoleucine Chemical compound CC[C@@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-UHNVWZDZSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- SNDPXSYFESPGGJ-UHFFFAOYSA-N L-norVal-OH Natural products CCCC(N)C(O)=O SNDPXSYFESPGGJ-UHFFFAOYSA-N 0.000 description 1
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-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
- 241000713666 Lentivirus Species 0.000 description 1
- 101000800648 Mus musculus DNA nucleotidylexotransferase Proteins 0.000 description 1
- 101100021581 Mus musculus Pol gene Proteins 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- OLNLSTNFRUFTLM-UHFFFAOYSA-N N-ethylasparagine Chemical compound CCNC(C(O)=O)CC(N)=O OLNLSTNFRUFTLM-UHFFFAOYSA-N 0.000 description 1
- YPIGGYHFMKJNKV-UHFFFAOYSA-N N-ethylglycine Chemical compound CC[NH2+]CC([O-])=O YPIGGYHFMKJNKV-UHFFFAOYSA-N 0.000 description 1
- 108010065338 N-ethylglycine Proteins 0.000 description 1
- AKCRVYNORCOYQT-YFKPBYRVSA-N N-methyl-L-valine Chemical compound CN[C@@H](C(C)C)C(O)=O AKCRVYNORCOYQT-YFKPBYRVSA-N 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 108020005091 Replication Origin Proteins 0.000 description 1
- 101100221606 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) COS7 gene Proteins 0.000 description 1
- 108010077895 Sarcosine Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 101710137500 T7 RNA polymerase Proteins 0.000 description 1
- 239000008051 TBE buffer Substances 0.000 description 1
- 101100388071 Thermococcus sp. (strain GE8) pol gene Proteins 0.000 description 1
- 102000006601 Thymidine Kinase Human genes 0.000 description 1
- 108020004440 Thymidine kinase Proteins 0.000 description 1
- 241001489220 Vanderwaltozyma polyspora Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 102100036976 X-ray repair cross-complementing protein 6 Human genes 0.000 description 1
- 101710124907 X-ray repair cross-complementing protein 6 Proteins 0.000 description 1
- 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 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 229940124277 aminobutyric acid Drugs 0.000 description 1
- 108010028263 bacteriophage T3 RNA polymerase Proteins 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 125000001314 canonical amino-acid group Chemical group 0.000 description 1
- 230000030570 cellular localization Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000005094 computer simulation Methods 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- YSMODUONRAFBET-UHFFFAOYSA-N delta-DL-hydroxylysine Natural products NCC(O)CCC(N)C(O)=O YSMODUONRAFBET-UHFFFAOYSA-N 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- YSMODUONRAFBET-UHNVWZDZSA-N erythro-5-hydroxy-L-lysine Chemical compound NC[C@H](O)CC[C@H](N)C(O)=O YSMODUONRAFBET-UHNVWZDZSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000012456 homogeneous solution Substances 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- QJHBJHUKURJDLG-UHFFFAOYSA-N hydroxy-L-lysine Natural products NCCCCC(NO)C(O)=O QJHBJHUKURJDLG-UHFFFAOYSA-N 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 239000006225 natural substrate Substances 0.000 description 1
- 230000030648 nucleus localization Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 238000002708 random mutagenesis Methods 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- YSMODUONRAFBET-WHFBIAKZSA-N threo-5-hydroxy-L-lysine Chemical compound NC[C@@H](O)CC[C@H](N)C(O)=O YSMODUONRAFBET-WHFBIAKZSA-N 0.000 description 1
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 239000004474 valine Substances 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
-
- 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)
- C12N9/1264—DNA nucleotidylexotransferase (2.7.7.31), i.e. terminal nucleotidyl transferase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/30—Nucleotides
- C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
-
- 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)
- C12Y207/07031—DNA nucleotidylexotransferase (2.7.7.31), i.e. terminal deoxynucleotidyl transferase
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR1655475A FR3052462A1 (fr) | 2016-06-14 | 2016-06-14 | Variants d'une adn polymerase de la famille polx |
| FR1655475 | 2016-06-14 | ||
| PCT/FR2017/051519 WO2017216472A2 (fr) | 2016-06-14 | 2017-06-13 | Variants d'une adn polymérase de la famille polx |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA3024184A1 true CA3024184A1 (en) | 2017-12-21 |
Family
ID=57348782
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA3024184A Pending CA3024184A1 (en) | 2016-06-14 | 2017-06-13 | Variants of a dna polymerase of the polx family |
Country Status (11)
| Country | Link |
|---|---|
| US (3) | US20200002690A1 (enExample) |
| EP (1) | EP3469075A2 (enExample) |
| JP (2) | JP7118510B2 (enExample) |
| KR (1) | KR102522263B1 (enExample) |
| CN (1) | CN109477080A (enExample) |
| AU (2) | AU2017286477C1 (enExample) |
| CA (1) | CA3024184A1 (enExample) |
| FR (2) | FR3052462A1 (enExample) |
| IL (1) | IL263503B2 (enExample) |
| SG (2) | SG10202009688QA (enExample) |
| WO (1) | WO2017216472A2 (enExample) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020099451A1 (en) * | 2018-11-14 | 2020-05-22 | Dna Script | Terminal deoxynucleotidyl transferase variants and uses thereof |
| WO2021116270A1 (en) * | 2019-12-12 | 2021-06-17 | Dna Script | Chimeric terminal deoxynucleotidyl transferases for template-free enzymatic synthesis of polynucleotides |
| JP2022513371A (ja) * | 2018-10-19 | 2022-02-07 | モレキュラー アッセンブリーズ， インコーポレイテッド | ポリデオキシヌクレオチド合成のための改変テンプレート非依存性酵素 |
| JP2023528477A (ja) * | 2020-06-03 | 2023-07-04 | モレキュラー アッセンブリーズ， インコーポレイテッド | ポリデオキシヌクレオチド合成のための改変された鋳型非依存的酵素 |
| EP4653525A1 (en) * | 2024-05-21 | 2025-11-26 | Uab "Biomatter Designs" | Engineered terminal deoxynucleotidyl transferase polymerases |
Families Citing this family (60)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018215803A1 (en) | 2017-05-26 | 2018-11-29 | Nuclera Nucleics Ltd | Use of terminal transferase enzyme in nucleic acid synthesis |
| EP3665274B1 (en) | 2017-08-07 | 2025-10-15 | DNA Script | Variants of family a dna polymerase and uses thereof |
| US10752887B2 (en) | 2018-01-08 | 2020-08-25 | Dna Script | Variants of terminal deoxynucleotidyl transferase and uses thereof |
| WO2019135007A1 (en) * | 2018-01-08 | 2019-07-11 | Dna Script | Variants of terminal deoxynucleotidyl transferase and uses thereof |
| US12421628B2 (en) | 2018-07-23 | 2025-09-23 | Dna Script | Massively parallel enzymatic synthesis of nucleic acid strands |
| KR20210104779A (ko) | 2018-12-13 | 2021-08-25 | 디엔에이 스크립트 | 세포 및 생체분자 상의 직접 올리고뉴클레오타이드 합성 |
| CN113272442B (zh) | 2019-01-03 | 2024-03-15 | Dna斯克瑞普特公司 | 寡核苷酸的集合的一锅合成 |
| GB201901501D0 (en) | 2019-02-04 | 2019-03-27 | Nuclera Nucleics Ltd | Modified terminal deoxynucleotidyl transferase (TdT) enzymes |
| KR20210125496A (ko) * | 2019-02-12 | 2021-10-18 | 디엔에이 스크립트 | 폴리뉴클레오티드의 주형-유리 효소 합성에서 효율적 생성물 절단 |
| ES2982551T3 (es) * | 2019-05-28 | 2024-10-16 | Dna Script | Variantes de desoxinucleotidil transferasa terminal y usos de las mismas |
| EP3744854A1 (en) * | 2019-05-28 | 2020-12-02 | DNA Script | Variants of terminal deoxynucleotidyl transferase and uses thereof |
| EP4004222A1 (en) | 2019-07-30 | 2022-06-01 | DNA Script | Template-free enzymatic synthesis of polynucleotides using poly(a) and poly(u) polymerases |
| US20220403435A1 (en) | 2019-08-01 | 2022-12-22 | Dna Script Sas | Increasing Long-Sequence Yields In Template-Free Enzymatic Synthesis of Polynucleotides |
| CN110331136B (zh) * | 2019-09-05 | 2019-12-24 | 中国科学院天津工业生物技术研究所 | 一种末端脱氧核糖核苷转移酶变体及其应用 |
| EP4028537A1 (en) | 2019-09-09 | 2022-07-20 | DNA Script | Template-free enzymatic polynucleotide synthesis using photocleavable linkages |
| JP2022549196A (ja) | 2019-09-23 | 2022-11-24 | ディーエヌエー スクリプト | ポリヌクレオチドの鋳型なし酵素的合成における長配列収率の増加 |
| JP2023501651A (ja) | 2019-11-13 | 2023-01-18 | ディーエヌエー スクリプト | ポリヌクレオチドの高効率テンプレートなしの酵素的合成 |
| WO2021122539A1 (en) | 2019-12-16 | 2021-06-24 | Dna Script | Template-free enzymatic polynucleotide synthesis using dismutationless terminal deoxynucleotidyl transferase variants |
| US20230089448A1 (en) | 2020-02-25 | 2023-03-23 | Dna Script | Method And Apparatus for Enzymatic Synthesis of Polynucleotides |
| KR20230002825A (ko) | 2020-04-20 | 2023-01-05 | 디엔에이 스크립트 | 말단 데옥시뉴클레오티딜 트랜스퍼라제 변이체 및 그의 용도 |
| US12110516B2 (en) | 2020-05-12 | 2024-10-08 | Illumina, Inc. | Thermostable terminal deoxynucleotidyl transferase |
| CN115916967A (zh) | 2020-05-12 | 2023-04-04 | 伊鲁米纳公司 | 使用重组末端脱氧核苷酸转移酶生成具有经修饰的碱基的核酸 |
| KR20230078992A (ko) | 2020-06-12 | 2023-06-05 | 신헬릭스 | 열안정성 효소를 사용한 핵산의 초기 주형 독립적 합성 |
| EP4164784B1 (en) | 2020-06-16 | 2025-06-25 | DNA Script | Method for enzymatic polynucleotide synthesis |
| EP4182472A1 (en) | 2020-07-15 | 2023-05-24 | DNA Script | Massively parallel enzymatic synthesis of polynucleotides |
| KR20230041975A (ko) | 2020-07-21 | 2023-03-27 | 일루미나, 싱가포르 피티이 엘티디 | Tdt-기반 효소적 핵산을 위한 기질로서의 염기-변형된 뉴클레오티드 |
| EP4192947A1 (en) * | 2020-08-04 | 2023-06-14 | Nuclera Nucleics Ltd | Modified terminal deoxynucleotidyl transferase (tdt) enzymes |
| GB2598152B (en) * | 2020-08-21 | 2025-04-16 | Nuclera Ltd | Modified terminal deoxynucleotidyl transferase (TdT) enzymes |
| WO2022063835A1 (en) | 2020-09-22 | 2022-03-31 | Dna Script | Stabilized n-terminally truncated terminal deoxynucleotidyl transferase variants and uses thereof |
| US20240052391A1 (en) | 2020-10-29 | 2024-02-15 | Dna Script | Enzymatic Synthesis of Polynucleotide Probes |
| US12391970B2 (en) | 2020-11-11 | 2025-08-19 | Microsoft Technology Licensing, Llc | Spatial control of polynucleotide synthesis by strand capping |
| CN112322715B (zh) * | 2020-11-17 | 2022-11-25 | 清华大学 | 一种核酸测序方法 |
| DE102022107811A1 (de) | 2021-04-02 | 2022-10-06 | Dna Script | Methoden und kits für die enzymatische synthese von zu-g4-neigenden polynukleotiden |
| DE102022114351A1 (de) | 2021-06-10 | 2022-12-15 | Dna Script | Enzymatische synthese von polynukleotiden mittels 3'-o-amino-2'-deoxyribonukleosid triphosphat monomeren |
| EP4392554A1 (en) | 2021-08-23 | 2024-07-03 | DNA Script | Method for generating double-stranded nucleic acid |
| US20240299945A1 (en) | 2021-09-03 | 2024-09-12 | Elegen Corporation | Multi-way bead-sorting devices, systems, and methods of use thereof using pressure sources |
| CA3236657A1 (en) | 2021-11-10 | 2023-05-19 | Daniel MCHALE | Novel terminal deoxynucleotidyl transferase (tdt) variant and uses thereof |
| WO2023083999A2 (en) | 2021-11-10 | 2023-05-19 | Dna Script | Novel terminal deoxynucleotidyl transferase (tdt) variants |
| CN116555216A (zh) * | 2022-01-28 | 2023-08-08 | 中国科学院天津工业生物技术研究所 | 可控合成单链dna的末端转移酶变体及应用 |
| WO2023170266A1 (en) | 2022-03-11 | 2023-09-14 | Dna Script | Automation station for enzymatic polynucleotide synthesis |
| WO2023170259A1 (en) | 2022-03-11 | 2023-09-14 | Dna Script | Modular accessory rack |
| WO2023170286A2 (en) | 2022-03-11 | 2023-09-14 | Dna Script | Alignment post and secure mechanism for enzymatic polynucleotide synthesis |
| WO2023170258A1 (en) | 2022-03-11 | 2023-09-14 | Dna Script | Apparatus for enzymatic synthesis of a plurality of polynucleotides comprising a condensation trap |
| WO2023240202A2 (en) * | 2022-06-08 | 2023-12-14 | Ansa Biotechnologies, Inc. | Modified terminal deoxynucleotidyl transferase polymerases |
| EP4555083A1 (en) | 2022-07-13 | 2025-05-21 | DNA Script | Method of purifying solution comprising polynucleotide |
| EP4630540A1 (en) | 2022-12-05 | 2025-10-15 | DNA Script | Variants of poly(a) polymerase and uses thereof |
| CN120359296A (zh) | 2022-12-09 | 2025-07-22 | Dna斯克瑞普特公司 | 体外酶促rna合成 |
| CN120603956A (zh) | 2022-12-31 | 2025-09-05 | Dna斯克瑞普特公司 | 用于酶试剂的喷墨递送的可变粘度油墨 |
| WO2024153643A1 (en) | 2023-01-16 | 2024-07-25 | Dna Script | Inkjet-assisted enzymatic nucleic acid synthesis |
| WO2024153642A1 (en) | 2023-01-16 | 2024-07-25 | Dna Script | Scarless template-free enzymatic synthesis of polynucleotides |
| EP4655411A2 (en) | 2023-01-26 | 2025-12-03 | DNA Script | Enzymatic synthesis of polynucleotide probes |
| CN121039286A (zh) | 2023-05-03 | 2025-11-28 | Dna斯克瑞普特公司 | 用于3'-o-氨基多核苷酸的去保护的试剂 |
| WO2024241324A1 (en) * | 2023-05-24 | 2024-11-28 | Scintium Ltd | Method for training an artificial intelligence (ai) model and generating a 3d computer-aided design (cad) and system therefor |
| WO2024256604A1 (en) | 2023-06-15 | 2024-12-19 | Dna Script | Salt addition during enzymatic polynucleotide synthesis |
| WO2025003434A1 (en) | 2023-06-30 | 2025-01-02 | Dna Script | Nucleic acid synthesis on reusable support |
| WO2025021984A1 (en) | 2023-07-27 | 2025-01-30 | Dna Script | Method for generating a plurality of polynucleotides |
| WO2025088024A1 (en) | 2023-10-24 | 2025-05-01 | Dna Script | Novel terminal deoxynucleotidyl transferase (tdt) variants and uses thereof |
| WO2025093659A1 (en) | 2023-11-04 | 2025-05-08 | Dna Script | 3'-o-aminooxymethyl blocked nucleotides and use thereof in enzymatic synthesis of polynucleotides |
| WO2025137872A1 (zh) * | 2023-12-26 | 2025-07-03 | 华大工程生物学长荡湖研究所 | 一种末端脱氧核糖核苷转移酶突变体及其制备方法和应用 |
| WO2025155239A1 (en) * | 2024-01-17 | 2025-07-24 | Agency For Science, Technology And Research | X family dna polymerases and uses thereof |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5004689A (en) | 1982-02-22 | 1991-04-02 | Biogen, Massachusetts | DNA sequences, recombinant DNA molecules and processes for producing human gamma interferon-like polypeptides in high yields |
| IL71691A (en) | 1984-04-27 | 1991-04-15 | Yeda Res & Dev | Production of interferon-ypsilon |
| US5047335A (en) | 1988-12-21 | 1991-09-10 | The Regents Of The University Of Calif. | Process for controlling intracellular glycosylation of proteins |
| BG52073B2 (en) | 1990-01-24 | 1996-04-30 | Inst Molekuljarna Biolog | Method for the preparation of recombinant human noncystein -interferon, free of n-end methionine |
| US6812339B1 (en) * | 2000-09-08 | 2004-11-02 | Applera Corporation | Polymorphisms in known genes associated with human disease, methods of detection and uses thereof |
| US9023633B2 (en) * | 2008-11-03 | 2015-05-05 | Kapa Biosystems | Chimeric DNA polymerases |
| CA2864394C (en) * | 2011-03-02 | 2021-10-19 | Jack Roth | A method of predicting a response to a tusc2 therapy |
| US10684289B2 (en) * | 2013-12-09 | 2020-06-16 | Adimab, Llc | Polyclonal mixtures of antibodies, and methods of making and using them |
| FR3020071B1 (fr) | 2014-04-17 | 2017-12-22 | Dna Script | Procede de synthese d'acides nucleiques, notamment d'acides nucleiques de grande longueur, utilisation du procede et kit pour la mise en œuvre du procede |
| FR3025201B1 (fr) | 2014-09-02 | 2018-10-12 | Dna Script | Nucleotides modifies pour la synthese d'acides nucleiques, un kit renfermant de tels nucleotides et leur utilisation pour la production de genes ou sequences d'acides nucleiques synthetiques |
| EP3699283A1 (en) * | 2014-10-20 | 2020-08-26 | Molecular Assemblies Inc. | Modified template-independent enzymes for polydeoxynucleotide systhesis |
-
2016
- 2016-06-14 FR FR1655475A patent/FR3052462A1/fr active Pending
-
2017
- 2017-06-13 KR KR1020187034921A patent/KR102522263B1/ko active Active
- 2017-06-13 SG SG10202009688QA patent/SG10202009688QA/en unknown
- 2017-06-13 IL IL263503A patent/IL263503B2/en unknown
- 2017-06-13 CN CN201780037427.0A patent/CN109477080A/zh active Pending
- 2017-06-13 US US16/309,414 patent/US20200002690A1/en not_active Abandoned
- 2017-06-13 SG SG11201809961TA patent/SG11201809961TA/en unknown
- 2017-06-13 JP JP2018564828A patent/JP7118510B2/ja active Active
- 2017-06-13 EP EP17746156.3A patent/EP3469075A2/fr active Pending
- 2017-06-13 WO PCT/FR2017/051519 patent/WO2017216472A2/fr not_active Ceased
- 2017-06-13 AU AU2017286477A patent/AU2017286477C1/en active Active
- 2017-06-13 CA CA3024184A patent/CA3024184A1/en active Pending
- 2017-07-07 FR FR1756408A patent/FR3053699B1/fr active Active
-
2022
- 2022-06-30 JP JP2022105854A patent/JP2022137127A/ja active Pending
- 2022-07-27 US US17/815,490 patent/US20230167421A1/en active Pending
-
2023
- 2023-11-21 US US18/516,616 patent/US20240124855A1/en active Pending
- 2023-12-13 AU AU2023282219A patent/AU2023282219A1/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2022513371A (ja) * | 2018-10-19 | 2022-02-07 | モレキュラー アッセンブリーズ， インコーポレイテッド | ポリデオキシヌクレオチド合成のための改変テンプレート非依存性酵素 |
| EP3867361A4 (en) * | 2018-10-19 | 2022-07-27 | Molecular Assemblies, Inc. | MODIFIED TEMPLATE-INDEPENDENT ENZYMES FOR POLYDEOXYNUCLEOTIDE SYNTHESIS |
| WO2020099451A1 (en) * | 2018-11-14 | 2020-05-22 | Dna Script | Terminal deoxynucleotidyl transferase variants and uses thereof |
| CN113302289A (zh) * | 2018-11-14 | 2021-08-24 | Dna斯克瑞普特公司 | 末端脱氧核苷酸转移酶变体及其用途 |
| US11859217B2 (en) | 2018-11-14 | 2024-01-02 | Dna Script | Terminal deoxynucleotidyl transferase variants and uses thereof |
| US12371680B2 (en) | 2018-11-14 | 2025-07-29 | Dna Script Sas | Terminal deoxynucleotidyl transferase variants and uses thereof |
| WO2021116270A1 (en) * | 2019-12-12 | 2021-06-17 | Dna Script | Chimeric terminal deoxynucleotidyl transferases for template-free enzymatic synthesis of polynucleotides |
| JP2023528477A (ja) * | 2020-06-03 | 2023-07-04 | モレキュラー アッセンブリーズ， インコーポレイテッド | ポリデオキシヌクレオチド合成のための改変された鋳型非依存的酵素 |
| EP4653525A1 (en) * | 2024-05-21 | 2025-11-26 | Uab "Biomatter Designs" | Engineered terminal deoxynucleotidyl transferase polymerases |
| WO2025243149A1 (en) * | 2024-05-21 | 2025-11-27 | Uab Biomatter Designs | Engineered terminal deoxynucleotidyl transferase polymerases |
Also Published As
| Publication number | Publication date |
|---|---|
| JP7118510B2 (ja) | 2022-08-16 |
| JP2019517799A (ja) | 2019-06-27 |
| WO2017216472A3 (fr) | 2018-03-15 |
| WO2017216472A2 (fr) | 2017-12-21 |
| IL263503B1 (en) | 2024-12-01 |
| SG11201809961TA (en) | 2018-12-28 |
| FR3053699A1 (fr) | 2018-01-12 |
| FR3052462A1 (fr) | 2017-12-15 |
| SG10202009688QA (en) | 2020-11-27 |
| EP3469075A2 (fr) | 2019-04-17 |
| JP2022137127A (ja) | 2022-09-21 |
| IL263503A (en) | 2019-02-03 |
| CN109477080A (zh) | 2019-03-15 |
| US20230167421A1 (en) | 2023-06-01 |
| AU2023282219A1 (en) | 2024-01-18 |
| KR102522263B1 (ko) | 2023-04-18 |
| IL263503B2 (en) | 2025-04-01 |
| US20240124855A1 (en) | 2024-04-18 |
| AU2017286477B2 (en) | 2023-09-14 |
| FR3053699B1 (fr) | 2021-04-02 |
| US20200002690A1 (en) | 2020-01-02 |
| KR20190024883A (ko) | 2019-03-08 |
| AU2017286477C1 (en) | 2023-12-21 |
| AU2017286477A1 (en) | 2018-12-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20230167421A1 (en) | Variants of a DNA Polymerase of the Polx Family | |
| JP7292442B2 (ja) | 改変アミノアシルｔＲＮＡ合成酵素およびその用途 | |
| CN113767168B (zh) | 高效引入赖氨酸衍生物的氨酰基—tRNA合成酶 | |
| CN112585264B (zh) | 重组型kod聚合酶 | |
| CN108239633B (zh) | 一种催化活性得到提高的d-阿洛酮糖-3-差向异构酶的突变体及其应用 | |
| CN107002074B (zh) | 活性型突变酶的制造方法及活性型突变酶、以及可溶性化突变蛋白的制造方法 | |
| AU2020242724B2 (en) | Aminoacyl-tRNA synthetase for efficiently introducing lysine derivative in protein | |
| CN109929820B (zh) | 新型的甘油单-二酰酯脂肪酶 | |
| JP2022534004A (ja) | 末端デオキシヌクレオチジルトランスフェラーゼの変異体及びその使用 | |
| CN109384852B (zh) | 重组Martentoxin的制备、表征及应用 | |
| CN109136209B (zh) | 肠激酶轻链突变体及其应用 | |
| CN102747060A (zh) | D-氨甲酰水解酶的突变体及其制备方法和应用 | |
| CN116334153A (zh) | 还原胺化酶及其制法和应用 | |
| RU2790662C1 (ru) | АМИНОАЦИЛ-тРНК-СИНТЕТАЗА, ЭФФЕКТИВНОЕ ВВЕДЕНИЕ ПРОИЗВОДНЫХ ЛИЗИНА | |
| CN100537754C (zh) | 一种s-腺苷甲硫氨酸生产菌株的构建及大规模发酵 | |
| CN105779415B (zh) | 一种新型的脂肪酶 | |
| CN105779414B (zh) | 一种新型的脂肪酶 | |
| CN105779417B (zh) | 一种新型的脂肪酶 | |
| CN105779416B (zh) | 一种新型的脂肪酶 | |
| WO2002070688A1 (fr) | Nouvelles proteines et adn codant pour elle | |
| CN112689674A (zh) | 葡聚糖亲和性标签及其应用 | |
| JP2005046093A (ja) | トランスポゾン転移酵素およびその利用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request |
Effective date: 20220608 |
|
| EEER | Examination request |
Effective date: 20220608 |
|
| EEER | Examination request |
Effective date: 20220608 |
|
| EEER | Examination request |
Effective date: 20220608 |
|
| EEER | Examination request |
Effective date: 20220608 |