JP4195815B2 - Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ（ａｇｔ）の使用方法 - Google Patents
Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ（ａｇｔ）の使用方法 Download PDFInfo
- Publication number
- JP4195815B2 JP4195815B2 JP2002582274A JP2002582274A JP4195815B2 JP 4195815 B2 JP4195815 B2 JP 4195815B2 JP 2002582274 A JP2002582274 A JP 2002582274A JP 2002582274 A JP2002582274 A JP 2002582274A JP 4195815 B2 JP4195815 B2 JP 4195815B2
- Authority
- JP
- Japan
- Prior art keywords
- fusion protein
- agt
- protein
- label
- substrate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 238000000034 method Methods 0.000 title claims abstract description 61
- 108010014722 Alkyl and Aryl Transferases Proteins 0.000 title claims abstract description 24
- 102000002226 Alkyl and Aryl Transferases Human genes 0.000 title claims abstract description 24
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 132
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 131
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 58
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 56
- 239000000758 substrate Substances 0.000 claims abstract description 51
- 238000001727 in vivo Methods 0.000 claims abstract description 19
- 239000000284 extract Substances 0.000 claims abstract description 12
- 235000018102 proteins Nutrition 0.000 claims description 53
- 238000002372 labelling Methods 0.000 claims description 37
- 239000000523 sample Substances 0.000 claims description 16
- 230000014509 gene expression Effects 0.000 claims description 14
- 238000000338 in vitro Methods 0.000 claims description 12
- 125000005647 linker group Chemical group 0.000 claims description 10
- 125000000217 alkyl group Chemical group 0.000 claims description 8
- 108091034117 Oligonucleotide Proteins 0.000 claims description 6
- 239000013604 expression vector Substances 0.000 claims description 6
- XWNJMSJGJFSGRY-UHFFFAOYSA-N 2-(benzylamino)-3,7-dihydropurin-6-one Chemical compound N1C=2N=CNC=2C(=O)N=C1NCC1=CC=CC=C1 XWNJMSJGJFSGRY-UHFFFAOYSA-N 0.000 claims description 5
- 229940124276 oligodeoxyribonucleotide Drugs 0.000 claims description 5
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims description 4
- 102000039446 nucleic acids Human genes 0.000 claims description 4
- 108020004707 nucleic acids Proteins 0.000 claims description 4
- 150000007523 nucleic acids Chemical class 0.000 claims description 4
- 239000002773 nucleotide Substances 0.000 claims description 4
- 125000003729 nucleotide group Chemical group 0.000 claims description 4
- 239000002202 Polyethylene glycol Substances 0.000 claims description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 3
- 239000002872 contrast media Substances 0.000 claims description 3
- 229920001223 polyethylene glycol Polymers 0.000 claims description 3
- 230000001131 transforming effect Effects 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 42
- 238000012546 transfer Methods 0.000 abstract description 10
- 238000001514 detection method Methods 0.000 abstract description 4
- 210000001723 extracellular space Anatomy 0.000 abstract description 3
- 230000001580 bacterial effect Effects 0.000 abstract description 2
- 238000004113 cell culture Methods 0.000 abstract 1
- 210000003527 eukaryotic cell Anatomy 0.000 abstract 1
- KRWMERLEINMZFT-UHFFFAOYSA-N O6-benzylguanine Chemical class C=12NC=NC2=NC(N)=NC=1OCC1=CC=CC=C1 KRWMERLEINMZFT-UHFFFAOYSA-N 0.000 description 18
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 14
- 108020004705 Codon Proteins 0.000 description 14
- 108091023040 Transcription factor Proteins 0.000 description 14
- 102000040945 Transcription factor Human genes 0.000 description 14
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 11
- 230000004927 fusion Effects 0.000 description 11
- 101710188306 Protein Y Proteins 0.000 description 10
- 150000001413 amino acids Chemical class 0.000 description 9
- 239000003446 ligand Substances 0.000 description 9
- 239000007787 solid Substances 0.000 description 9
- -1 alkyl guanine Chemical compound 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 150000003384 small molecules Chemical class 0.000 description 8
- 239000013076 target substance Substances 0.000 description 8
- 108010090804 Streptavidin Proteins 0.000 description 7
- 230000004913 activation Effects 0.000 description 7
- 229960002685 biotin Drugs 0.000 description 7
- 235000020958 biotin Nutrition 0.000 description 7
- 239000011616 biotin Substances 0.000 description 7
- 229940079593 drug Drugs 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 239000002953 phosphate buffered saline Substances 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 6
- 241000588724 Escherichia coli Species 0.000 description 6
- 230000027455 binding Effects 0.000 description 6
- 230000004568 DNA-binding Effects 0.000 description 5
- 102100024746 Dihydrofolate reductase Human genes 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 5
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 5
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 5
- 102000003992 Peroxidases Human genes 0.000 description 5
- 108020001096 dihydrofolate reductase Proteins 0.000 description 5
- 239000005090 green fluorescent protein Substances 0.000 description 5
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 5
- 230000003993 interaction Effects 0.000 description 5
- 108040007629 peroxidase activity proteins Proteins 0.000 description 5
- 230000009870 specific binding Effects 0.000 description 5
- 238000001262 western blot Methods 0.000 description 5
- 210000005253 yeast cell Anatomy 0.000 description 5
- 108090001008 Avidin Proteins 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 150000003254 radicals Chemical class 0.000 description 4
- 101100327917 Caenorhabditis elegans chup-1 gene Proteins 0.000 description 3
- 108020004414 DNA Proteins 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
- 229940072107 ascorbate Drugs 0.000 description 3
- 235000010323 ascorbic acid Nutrition 0.000 description 3
- 239000011668 ascorbic acid Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 108091006004 biotinylated proteins Proteins 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 239000013522 chelant Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000002512 chemotherapy Methods 0.000 description 3
- 230000001351 cycling effect Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 230000002934 lysing effect Effects 0.000 description 3
- 229930182817 methionine Natural products 0.000 description 3
- 210000004940 nucleus Anatomy 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 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 2
- 238000002965 ELISA Methods 0.000 description 2
- 238000012286 ELISA Assay Methods 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 101100385969 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) CYC8 gene Proteins 0.000 description 2
- 101100276456 Schizosaccharomyces pombe (strain 972 / ATCC 24843) ssn6 gene Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
- 230000006287 biotinylation Effects 0.000 description 2
- 238000007413 biotinylation Methods 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 230000005284 excitation Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 210000005260 human cell Anatomy 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- FDZZZRQASAIRJF-UHFFFAOYSA-M malachite green Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](C)C)C=C1 FDZZZRQASAIRJF-UHFFFAOYSA-M 0.000 description 2
- 229940107698 malachite green Drugs 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- PXQPEWDEAKTCGB-UHFFFAOYSA-N orotic acid Chemical compound OC(=O)C1=CC(=O)NC(=O)N1 PXQPEWDEAKTCGB-UHFFFAOYSA-N 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000003498 protein array Methods 0.000 description 2
- 230000012743 protein tagging Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 1
- 150000003923 2,5-pyrrolediones Chemical class 0.000 description 1
- 102100033647 Activity-regulated cytoskeleton-associated protein Human genes 0.000 description 1
- 102000011724 DNA Repair Enzymes Human genes 0.000 description 1
- 108010076525 DNA Repair Enzymes Proteins 0.000 description 1
- 230000007118 DNA alkylation Effects 0.000 description 1
- 108010078339 DNA alkyltransferase Proteins 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 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 1
- 241000282412 Homo Species 0.000 description 1
- 101710128836 Large T antigen Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 101000966481 Mus musculus Dihydrofolate reductase Proteins 0.000 description 1
- ZRKWMRDKSOPRRS-UHFFFAOYSA-N N-Methyl-N-nitrosourea Chemical compound O=NN(C)C(N)=O ZRKWMRDKSOPRRS-UHFFFAOYSA-N 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 241000907663 Siproeta stelenes Species 0.000 description 1
- 102000018679 Tacrolimus Binding Proteins Human genes 0.000 description 1
- 108010027179 Tacrolimus Binding Proteins Proteins 0.000 description 1
- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 150000001934 cyclohexanes Chemical class 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 229960003901 dacarbazine Drugs 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000012039 electrophile Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 235000021384 green leafy vegetables Nutrition 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 108091006058 immobilized fusion proteins Proteins 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 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 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000030648 nucleus localization Effects 0.000 description 1
- 229960005010 orotic acid Drugs 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000023603 positive regulation of transcription initiation, DNA-dependent Effects 0.000 description 1
- CPTBDICYNRMXFX-UHFFFAOYSA-N procarbazine Chemical compound CNNCC1=CC=C(C(=O)NC(C)C)C=C1 CPTBDICYNRMXFX-UHFFFAOYSA-N 0.000 description 1
- 229960000624 procarbazine Drugs 0.000 description 1
- 238000000734 protein sequencing Methods 0.000 description 1
- 230000004850 protein–protein interaction Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 125000006853 reporter group Chemical group 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229960004964 temozolomide Drugs 0.000 description 1
- 238000010399 three-hybrid screening Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 108091006107 transcriptional repressors Proteins 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000000539 two dimensional gel electrophoresis Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- 238000001086 yeast two-hybrid system Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Enzymes And Modification Thereof (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US28276601P | 2001-04-10 | 2001-04-10 | |
| PCT/GB2002/001636 WO2002083937A2 (en) | 2001-04-10 | 2002-04-05 | Methods using o6-alkylguanine-dna alkyltransferases |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2007328757A Division JP4226053B2 (ja) | 2001-04-10 | 2007-12-20 | Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ（ａｇｔ）の使用方法 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2004532028A JP2004532028A (ja) | 2004-10-21 |
| JP2004532028A5 JP2004532028A5 (https=) | 2008-10-02 |
| JP4195815B2 true JP4195815B2 (ja) | 2008-12-17 |
Family
ID=23083030
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2002582274A Expired - Lifetime JP4195815B2 (ja) | 2001-04-10 | 2002-04-05 | Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ（ａｇｔ）の使用方法 |
| JP2007328757A Expired - Fee Related JP4226053B2 (ja) | 2001-04-10 | 2007-12-20 | Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ（ａｇｔ）の使用方法 |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2007328757A Expired - Fee Related JP4226053B2 (ja) | 2001-04-10 | 2007-12-20 | Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ（ａｇｔ）の使用方法 |
Country Status (14)
| Country | Link |
|---|---|
| US (2) | US7939284B2 (https=) |
| EP (4) | EP1410023B1 (https=) |
| JP (2) | JP4195815B2 (https=) |
| KR (1) | KR20040039194A (https=) |
| CN (2) | CN1295510C (https=) |
| AT (3) | ATE419378T1 (https=) |
| AU (1) | AU2002251257B2 (https=) |
| CA (1) | CA2443570A1 (https=) |
| DE (3) | DE60212642T2 (https=) |
| DK (1) | DK1410023T3 (https=) |
| IL (1) | IL157880A0 (https=) |
| NZ (3) | NZ528594A (https=) |
| WO (1) | WO2002083937A2 (https=) |
| ZA (1) | ZA200307442B (https=) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006501286A (ja) * | 2002-10-03 | 2006-01-12 | エコル・ポリテクニック・フェデラル・ドゥ・ローザンヌ（エーペーエフエル） | Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ |
Families Citing this family (53)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050062588A (ko) * | 2002-10-03 | 2005-06-23 | 에콜 폴리테크니크 페데랄레 드 로잔 | Ｏ6-알킬구아닌-ｄｎａ 알킬트랜스퍼라제용 기질 |
| EP1594962B1 (en) | 2003-01-31 | 2014-08-27 | Promega Corporation | Covalent tethering of functional groups to proteins |
| US7429472B2 (en) | 2003-01-31 | 2008-09-30 | Promega Corporation | Method of immobilizing a protein or molecule via a mutant dehalogenase that is bound to an immobilized dehalogenase substrate and linked directly or indirectly to the protein or molecule |
| AU2004242249A1 (en) * | 2003-05-23 | 2004-12-02 | Epfl-Ecole Polytechnique Federale De Lausanne | Methods for protein labeling based on acyl carrier protein |
| US8163479B2 (en) | 2004-03-02 | 2012-04-24 | Ecole Polytechnique Federale De Lausanne | Specific substrates for O6-alkylguanine-DNA alkyltransferase |
| US7888090B2 (en) | 2004-03-02 | 2011-02-15 | Ecole Polytechnique Federale De Lausanne | Mutants of O6-alkylguanine-DNA alkyltransferase |
| US7425436B2 (en) | 2004-07-30 | 2008-09-16 | Promega Corporation | Covalent tethering of functional groups to proteins and substrates therefor |
| US7825096B2 (en) | 2004-09-08 | 2010-11-02 | The United States Of America As Represented By The Department Of Health And Human Services | O6-alkylguanine-DNA alkyltransferase inactivators and beta-glucuronidase cleavable prodrugs |
| US7273714B2 (en) * | 2005-03-11 | 2007-09-25 | Vion Pharmaceuticals, Inc. | Alkylguanyltransferase assays |
| WO2006114409A1 (en) * | 2005-04-27 | 2006-11-02 | Covalys Biosciences Ag | Pyrimidines reacting with o6-alkylguanine-dna alkyltransferase |
| FR2890174B1 (fr) | 2005-08-30 | 2009-04-24 | Cis Bio Internat Sa | Procede pour la mise en evidence d'un processus biologique par mesure d'un fret |
| FR2890446B1 (fr) | 2005-09-05 | 2008-04-18 | Cis Bio Internat Sa | Methode de detection d'interaction intracellulaire entre bio-molecules |
| CA2656192C (en) | 2006-06-30 | 2016-05-03 | Ambit Biosciences Corporation | Detectable nucleic acid tag |
| EP2374875B1 (en) | 2006-10-30 | 2015-09-02 | Promega Corporation | Mutant hydrolase proteins with enhanced kinetics and funcitional expression |
| US7897737B2 (en) | 2006-12-05 | 2011-03-01 | Lasergen, Inc. | 3′-OH unblocked, nucleotides and nucleosides, base modified with photocleavable, terminating groups and methods for their use in DNA sequencing |
| MX2009005936A (es) * | 2006-12-05 | 2010-01-15 | Lasergen Inc | Nucleotidos y nucleosidos marcados de foto-escision y nucleotidos y nucleosidos marcados y metodos para su uso en secunciacion de adn. |
| EP1978093A1 (en) * | 2007-04-04 | 2008-10-08 | Ecole Polytechnique Fédérale de Lausanne (EPFL) | Method of crosslinking two objects of interest |
| GB0711560D0 (en) * | 2007-06-14 | 2007-07-25 | Innova Biosciences Ltd | Improvements in or relating to production of conjugates |
| WO2009058814A1 (en) | 2007-10-29 | 2009-05-07 | New England Biolabs, Inc. | Methods and compositions for detection and enrichment of target small rnas |
| EP2093564A1 (en) | 2008-02-25 | 2009-08-26 | Technische Universität Dresden Medizinische Fakultät Carl Gustav Carus | Method for distinguishing secretory granules of different ages |
| KR101677772B1 (ko) | 2008-06-11 | 2016-11-18 | 레이저젠, 인코퍼레이티드 | 뉴클레오타이드 및 뉴클레오사이드 그리고 시퀀싱에서의 이들의 이용 방법 |
| FR2936245B1 (fr) * | 2008-09-23 | 2012-07-06 | Cis Bio Int | Nouveaux substrats d'o6-alkylguanine-adn alkyltransferase et ses mutants. |
| DE112011100297B4 (de) | 2010-01-20 | 2017-08-24 | New England Biolabs, Inc. | Zusammensetzungen, Verfahren und damit in Zusammenhang stehende Anwendungen zum Spalten modifizierter DNA |
| WO2012037361A2 (en) | 2010-09-15 | 2012-03-22 | Endacea, Inc. | Methods of use and kit for measurement of lipopolysaccharide with a time resolved fluorescence based assay |
| MX342195B (es) | 2011-09-13 | 2016-09-20 | Lasergen Inc | Nucleótidos de terminación de rápida fotodescomposición 5-metoxi, 3' -oh no bloqueados y métodos para secuenciación de ácido nucleico. |
| MX349562B (es) * | 2011-12-09 | 2017-08-02 | Pasteur Institut | Ensayo de inmuno deteccion multiplex. |
| US9200260B2 (en) | 2012-03-15 | 2015-12-01 | New England Biolabs, Inc. | Compositions and methods for the transfer of a hexosamine to a modified nucleotide in a nucleic acid |
| JP6441893B2 (ja) | 2013-03-19 | 2018-12-19 | ディレクティド・ジェノミクス・エル・エル・シー | 標的配列の濃縮 |
| JP5781589B2 (ja) * | 2013-12-20 | 2015-09-24 | レーザーゲン インコーポレイテッド | 光開裂性標識ヌクレオチドとヌクレオシドおよび標識ヌクレオチドとヌクレオシドならびにｄｎａ配列決定におけるそれらの使用法 |
| WO2016003795A1 (en) | 2014-07-02 | 2016-01-07 | New England Biolabs, Inc. | Universal n-glycan binding reagent |
| JP2017537657A (ja) | 2014-12-11 | 2017-12-21 | ニユー・イングランド・バイオレイブス・インコーポレイテツド | 標的配列の濃縮 |
| CN104844710B (zh) * | 2015-04-13 | 2018-02-02 | 西北大学 | 定向固定化pega复合型树酯的制备方法及应用 |
| CN109661407A (zh) | 2016-06-06 | 2019-04-19 | 西北大学 | 融合蛋白构建体 |
| CN113277985B (zh) | 2016-07-20 | 2023-07-28 | 纳莹(上海)生物科技有限公司 | 一种荧光探针及其制备方法和用途 |
| US10934533B1 (en) | 2018-10-10 | 2021-03-02 | New England Biolabs, Inc. | Variant DNA polymerases having improved properties and method for improved isothermal amplification of a target DNA |
| CN111253417B (zh) * | 2019-09-04 | 2022-03-25 | 大连民族大学 | 一种嘌呤衍生物的合成及应用 |
| JP7727628B2 (ja) | 2019-12-03 | 2025-08-21 | アラマー バイオサイエンシーズ， インク． | 核酸結合免疫サンドイッチアッセイ（ｎｕｌｉｓａ） |
| EP4103701A1 (en) | 2020-02-13 | 2022-12-21 | New England Biolabs, Inc. | Variant family d dna polymerases |
| WO2021163559A1 (en) | 2020-02-13 | 2021-08-19 | New England Biolabs, Inc. | Variant family d dna polymerases |
| WO2021183921A1 (en) | 2020-03-12 | 2021-09-16 | New England Biolabs, Inc. | A rapid diagnostic test for lamp |
| CN112010983A (zh) * | 2020-08-11 | 2020-12-01 | 西北大学 | 止咳平喘中药活性成分靶向筛选方法及试剂盒 |
| WO2022040443A2 (en) | 2020-08-21 | 2022-02-24 | New England Biolabs, Inc. | A rapid diagnostic test for lamp |
| US11993792B2 (en) | 2021-05-27 | 2024-05-28 | New England Biolabs, Inc. | DNase I variants, compositions, methods, and kits |
| WO2022250676A1 (en) | 2021-05-27 | 2022-12-01 | New England Biolabs, Inc. | Dnase i variants, compositions, methods, and kits |
| CA3236352A1 (en) | 2021-11-24 | 2023-06-01 | Romualdas Vaisvila | Double-stranded dna deaminases |
| WO2023197015A1 (en) | 2022-04-08 | 2023-10-12 | New England Biolabs, Inc. | Compositions and analysis of dephosphorylated oligoribonucleotides |
| EP4658673A1 (en) | 2023-02-01 | 2025-12-10 | New England Biolabs, Inc. | Duplex-specific dnases |
| US20250115953A1 (en) | 2023-10-05 | 2025-04-10 | New England Biolabs, Inc. | Methylcytosine-Selective Deaminases and Uses Thereof |
| WO2025129193A1 (en) | 2023-12-15 | 2025-06-19 | New England Biolabs, Inc. | Glycosyltransferases and related methods |
| WO2025137453A1 (en) | 2023-12-20 | 2025-06-26 | New England Biolabs, Inc. | Compositions, methods, kits, and instruments for analyzing rna structure |
| GB202400129D0 (en) | 2024-01-04 | 2024-02-21 | Cambridge Entpr Ltd | Nucleic acid barcodes |
| WO2025224010A1 (en) | 2024-04-24 | 2025-10-30 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e. V. | Improved o6-alkylguanine-dna alkyltransferase substrates |
| WO2025227034A1 (en) | 2024-04-26 | 2025-10-30 | New England Biolabs, Inc. | Salt active nuclease compositions and methods |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9224888D0 (en) * | 1992-11-27 | 1993-01-13 | Univ Singapore | Monoclonal antibody |
| DK0702683T3 (da) * | 1993-06-08 | 2004-02-02 | Cancer Rec Tech Ltd | 06-substituerede guaninderivater, fremgangsmåde til fremstilling deraf og anvendelse deraf i behandling af tumorceller |
| DE19903895A1 (de) | 1999-02-01 | 2000-08-03 | Kai Johnsson | Selektive Immobilisierung und Nachweis der O·6·-Alkylguanin-DNA-Alkyltransferase in der aktiven Form |
-
2002
- 2002-04-05 AT AT06111733T patent/ATE419378T1/de not_active IP Right Cessation
- 2002-04-05 US US10/474,796 patent/US7939284B2/en not_active Expired - Lifetime
- 2002-04-05 CN CNB028081137A patent/CN1295510C/zh not_active Expired - Lifetime
- 2002-04-05 DE DE60212642T patent/DE60212642T2/de not_active Expired - Lifetime
- 2002-04-05 EP EP02720187A patent/EP1410023B1/en not_active Expired - Lifetime
- 2002-04-05 EP EP06111733A patent/EP1696234B1/en not_active Expired - Lifetime
- 2002-04-05 JP JP2002582274A patent/JP4195815B2/ja not_active Expired - Lifetime
- 2002-04-05 DE DE60230689T patent/DE60230689D1/de not_active Expired - Lifetime
- 2002-04-05 AU AU2002251257A patent/AU2002251257B2/en not_active Ceased
- 2002-04-05 WO PCT/GB2002/001636 patent/WO2002083937A2/en not_active Ceased
- 2002-04-05 AT AT02720187T patent/ATE331220T1/de not_active IP Right Cessation
- 2002-04-05 CA CA002443570A patent/CA2443570A1/en not_active Abandoned
- 2002-04-05 KR KR10-2003-7012899A patent/KR20040039194A/ko not_active Abandoned
- 2002-04-05 EP EP08168182A patent/EP2037271B1/en not_active Expired - Lifetime
- 2002-04-05 CN CN2006101603317A patent/CN1975422B/zh not_active Expired - Lifetime
- 2002-04-05 NZ NZ528594A patent/NZ528594A/en unknown
- 2002-04-05 NZ NZ537939A patent/NZ537939A/en unknown
- 2002-04-05 EP EP10075132.0A patent/EP2211177B1/en not_active Expired - Lifetime
- 2002-04-05 NZ NZ540410A patent/NZ540410A/en unknown
- 2002-04-05 IL IL15788002A patent/IL157880A0/xx not_active IP Right Cessation
- 2002-04-05 DK DK02720187T patent/DK1410023T3/da active
- 2002-04-05 AT AT08168182T patent/ATE482395T1/de not_active IP Right Cessation
- 2002-04-05 DE DE60237792T patent/DE60237792D1/de not_active Expired - Lifetime
-
2003
- 2003-09-25 ZA ZA200307442A patent/ZA200307442B/xx unknown
-
2007
- 2007-12-20 JP JP2007328757A patent/JP4226053B2/ja not_active Expired - Fee Related
-
2011
- 2011-01-24 US US13/012,234 patent/US8367361B2/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006501286A (ja) * | 2002-10-03 | 2006-01-12 | エコル・ポリテクニック・フェデラル・ドゥ・ローザンヌ（エーペーエフエル） | Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ |
Also Published As
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4195815B2 (ja) | Ｏ６−アルキルグアニン−ｄｎａアルキルトランスフェラーゼ（ａｇｔ）の使用方法 | |
| JP2004532028A5 (https=) | ||
| AU2002251257A1 (en) | Methods of Using O6-alkylguanine-DNA Alkyltransferases | |
| JP7774084B2 (ja) | 多分子ルシフェラーゼ | |
| US7666612B2 (en) | Methods for protein labeling based on acyl carrier protein | |
| JP2025162556A (ja) | 多分子ルシフェラーゼペプチド及びポリペプチド | |
| US20060292651A1 (en) | Protein labelling with oº-alkylguanine-dna alkyltrnsferase | |
| Ozawa et al. | A minimal peptide sequence that targets fluorescent and functional proteins into the mitochondrial intermembrane space | |
| HK1068406B (en) | Methods using o6 - alkylguanine - dna alkyltransferases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20050328 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20070925 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20071220 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20080805 |
|
| A524 | Written submission of copy of amendment under article 19 pct |
Free format text: JAPANESE INTERMEDIATE CODE: A524 Effective date: 20080808 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20080909 |
|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20080929 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 4195815 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20111003 Year of fee payment: 3 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20121003 Year of fee payment: 4 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20131003 Year of fee payment: 5 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| EXPY | Cancellation because of completion of term |