JP2005512578A - Ｐｃｒをベースとする高効率のポリペプチドスクリーニング - Google Patents
Ｐｃｒをベースとする高効率のポリペプチドスクリーニング Download PDFInfo
- Publication number
- JP2005512578A JP2005512578A JP2003554934A JP2003554934A JP2005512578A JP 2005512578 A JP2005512578 A JP 2005512578A JP 2003554934 A JP2003554934 A JP 2003554934A JP 2003554934 A JP2003554934 A JP 2003554934A JP 2005512578 A JP2005512578 A JP 2005512578A
- Authority
- JP
- Japan
- Prior art keywords
- template
- amplification
- reaction
- replicable
- pcr
- 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
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 22
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 22
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 22
- 238000012216 screening Methods 0.000 title claims abstract description 21
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 57
- 230000003321 amplification Effects 0.000 claims abstract description 46
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 46
- 238000002703 mutagenesis Methods 0.000 claims abstract description 28
- 231100000350 mutagenesis Toxicity 0.000 claims abstract description 28
- 238000013519 translation Methods 0.000 claims abstract description 27
- 102000004190 Enzymes Human genes 0.000 claims abstract description 23
- 108090000790 Enzymes Proteins 0.000 claims abstract description 23
- 238000000338 in vitro Methods 0.000 claims abstract description 21
- 238000013518 transcription Methods 0.000 claims abstract description 21
- 230000035897 transcription Effects 0.000 claims abstract description 21
- 230000010076 replication Effects 0.000 claims abstract description 13
- 238000006243 chemical reaction Methods 0.000 claims description 66
- 238000000034 method Methods 0.000 claims description 65
- 239000000047 product Substances 0.000 claims description 53
- 210000004027 cell Anatomy 0.000 claims description 29
- 239000003153 chemical reaction reagent Substances 0.000 claims description 16
- 230000014509 gene expression Effects 0.000 claims description 15
- 125000003729 nucleotide group Chemical group 0.000 claims description 14
- 230000006798 recombination Effects 0.000 claims description 13
- 238000005215 recombination Methods 0.000 claims description 13
- 238000010367 cloning Methods 0.000 claims description 11
- 239000002773 nucleotide Substances 0.000 claims description 10
- 108700026244 Open Reading Frames Proteins 0.000 claims description 8
- 239000011541 reaction mixture Substances 0.000 claims description 7
- 230000009466 transformation Effects 0.000 claims description 7
- 239000012634 fragment Substances 0.000 claims description 6
- 238000011144 upstream manufacturing Methods 0.000 claims description 6
- 206010059866 Drug resistance Diseases 0.000 claims description 5
- 239000007795 chemical reaction product Substances 0.000 claims description 4
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 210000004102 animal cell Anatomy 0.000 claims description 2
- 239000000427 antigen Substances 0.000 claims description 2
- 102000036639 antigens Human genes 0.000 claims description 2
- 108091007433 antigens Proteins 0.000 claims description 2
- 230000002538 fungal effect Effects 0.000 claims description 2
- 244000052769 pathogen Species 0.000 claims description 2
- 230000005030 transcription termination Effects 0.000 claims description 2
- 230000001717 pathogenic effect Effects 0.000 claims 1
- 230000001131 transforming effect Effects 0.000 claims 1
- 108091033319 polynucleotide Proteins 0.000 abstract description 34
- 102000040430 polynucleotide Human genes 0.000 abstract description 34
- 239000002157 polynucleotide Substances 0.000 abstract description 34
- 230000000694 effects Effects 0.000 abstract description 12
- 108091026890 Coding region Proteins 0.000 abstract description 9
- 239000000758 substrate Substances 0.000 abstract description 4
- 102000004169 proteins and genes Human genes 0.000 description 37
- 235000018102 proteins Nutrition 0.000 description 36
- 230000014616 translation Effects 0.000 description 31
- 108020004414 DNA Proteins 0.000 description 21
- 230000035772 mutation Effects 0.000 description 17
- 238000003786 synthesis reaction Methods 0.000 description 15
- 230000015572 biosynthetic process Effects 0.000 description 12
- 239000000284 extract Substances 0.000 description 12
- 239000000203 mixture Substances 0.000 description 10
- 150000001413 amino acids Chemical group 0.000 description 9
- 239000000872 buffer Substances 0.000 description 9
- 102000039446 nucleic acids Human genes 0.000 description 9
- 108020004707 nucleic acids Proteins 0.000 description 9
- 150000007523 nucleic acids Chemical class 0.000 description 9
- 235000001014 amino acid Nutrition 0.000 description 7
- 238000001243 protein synthesis Methods 0.000 description 7
- 239000013598 vector Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 5
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 5
- 108091008146 restriction endonucleases Proteins 0.000 description 5
- 239000007790 solid phase Substances 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000003362 replicative effect Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 4
- PFNFFQXMRSDOHW-UHFFFAOYSA-N spermine Chemical compound NCCCNCCCCNCCCN PFNFFQXMRSDOHW-UHFFFAOYSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical class CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 108091093088 Amplicon Proteins 0.000 description 3
- 108020004705 Codon Proteins 0.000 description 3
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 3
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 239000005090 green fluorescent protein Substances 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000007857 nested PCR Methods 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000376 reactant Substances 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 125000000022 2-aminoethyl group Chemical group [H]C([*])([H])C([H])([H])N([H])[H] 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010067770 Endopeptidase K Proteins 0.000 description 2
- 108060002716 Exonuclease Proteins 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 102000018120 Recombinases Human genes 0.000 description 2
- 108010091086 Recombinases Proteins 0.000 description 2
- 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 2
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical compound C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 210000004671 cell-free system Anatomy 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000012790 confirmation Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 102000013165 exonuclease Human genes 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 229920000642 polymer Chemical class 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 239000011535 reaction buffer Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 229920002477 rna polymer Polymers 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 238000002741 site-directed mutagenesis Methods 0.000 description 2
- 229940063673 spermidine Drugs 0.000 description 2
- 229940063675 spermine Drugs 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000001226 triphosphate Substances 0.000 description 2
- 235000011178 triphosphate Nutrition 0.000 description 2
- AUFGTPPARQZWDO-YUZLPWPTSA-N 10-formyltetrahydrofolate Chemical compound C1NC=2NC(N)=NC(=O)C=2NC1CN(C=O)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 AUFGTPPARQZWDO-YUZLPWPTSA-N 0.000 description 1
- MWBWWFOAEOYUST-UHFFFAOYSA-N 2-aminopurine Chemical compound NC1=NC=C2N=CNC2=N1 MWBWWFOAEOYUST-UHFFFAOYSA-N 0.000 description 1
- WTLKTXIHIHFSGU-UHFFFAOYSA-N 2-nitrosoguanidine Chemical compound NC(N)=NN=O WTLKTXIHIHFSGU-UHFFFAOYSA-N 0.000 description 1
- KKAJSJJFBSOMGS-UHFFFAOYSA-N 3,6-diamino-10-methylacridinium chloride Chemical compound [Cl-].C1=C(N)C=C2[N+](C)=C(C=C(N)C=C3)C3=CC2=C1 KKAJSJJFBSOMGS-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- LQLQRFGHAALLLE-UHFFFAOYSA-N 5-bromouracil Chemical compound BrC1=CNC(=O)NC1=O LQLQRFGHAALLLE-UHFFFAOYSA-N 0.000 description 1
- 102000052866 Amino Acyl-tRNA Synthetases Human genes 0.000 description 1
- 108700028939 Amino Acyl-tRNA Synthetases Proteins 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 108010035563 Chloramphenicol O-acetyltransferase Proteins 0.000 description 1
- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 102100021309 Elongation factor Ts, mitochondrial Human genes 0.000 description 1
- 102100033238 Elongation factor Tu, mitochondrial Human genes 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 1
- -1 IF-2 and IF-3 Proteins 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 102000005431 Molecular Chaperones Human genes 0.000 description 1
- 108010006519 Molecular Chaperones Proteins 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 102000010562 Peptide Elongation Factor G Human genes 0.000 description 1
- 108010077742 Peptide Elongation Factor G Proteins 0.000 description 1
- 108010049977 Peptide Elongation Factor Tu Proteins 0.000 description 1
- 108010044843 Peptide Initiation Factors Proteins 0.000 description 1
- 102000005877 Peptide Initiation Factors Human genes 0.000 description 1
- 108090000279 Peptidyltransferases Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- WDVSHHCDHLJJJR-UHFFFAOYSA-N Proflavine Chemical compound C1=CC(N)=CC2=NC3=CC(N)=CC=C3C=C21 WDVSHHCDHLJJJR-UHFFFAOYSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 108091081062 Repeated sequence (DNA) Proteins 0.000 description 1
- 101100221606 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) COS7 gene Proteins 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical class [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- RYYWUUFWQRZTIU-UHFFFAOYSA-N Thiophosphoric acid Chemical class OP(O)(S)=O RYYWUUFWQRZTIU-UHFFFAOYSA-N 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- IVOMOUWHDPKRLL-UHFFFAOYSA-N UNPD107823 Natural products O1C2COP(O)(=O)OC2C(O)C1N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-UHFFFAOYSA-N 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- MZVQCMJNVPIDEA-UHFFFAOYSA-N [CH2]CN(CC)CC Chemical group [CH2]CN(CC)CC MZVQCMJNVPIDEA-UHFFFAOYSA-N 0.000 description 1
- 229940023020 acriflavine Drugs 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 238000000376 autoradiography Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000001311 chemical methods and process Methods 0.000 description 1
- 239000002962 chemical mutagen Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 229940095074 cyclic amp Drugs 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 230000009615 deamination Effects 0.000 description 1
- 238000006481 deamination reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000013024 dilution buffer Substances 0.000 description 1
- NAGJZTKCGNOGPW-UHFFFAOYSA-N dithiophosphoric acid Chemical class OP(O)(S)=S NAGJZTKCGNOGPW-UHFFFAOYSA-N 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 108010063460 elongation factor T Proteins 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 229930192878 garvin Natural products 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 229920001519 homopolymer Polymers 0.000 description 1
- 244000052637 human pathogen Species 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 239000000138 intercalating agent Substances 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 150000002696 manganese Chemical class 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 229960000901 mepacrine Drugs 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229960004452 methionine Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000001293 nucleolytic effect Effects 0.000 description 1
- 238000004091 panning Methods 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 150000008300 phosphoramidites Chemical class 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 230000001323 posttranslational effect Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 229960000286 proflavine Drugs 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- GPKJTRJOBQGKQK-UHFFFAOYSA-N quinacrine Chemical compound C1=C(OC)C=C2C(NC(C)CCCN(CC)CC)=C(C=CC(Cl)=C3)C3=NC2=C1 GPKJTRJOBQGKQK-UHFFFAOYSA-N 0.000 description 1
- 238000002708 random mutagenesis Methods 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 210000001995 reticulocyte Anatomy 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 108091006106 transcriptional activators Proteins 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 125000002264 triphosphate group Chemical class [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 1
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
- 150000003952 β-lactams Chemical class 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
- 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
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1086—Preparation or screening of expression libraries, e.g. reporter assays
-
- 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
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Crystallography & Structural Chemistry (AREA)
- Bioinformatics & Computational Biology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US34197801P | 2001-12-19 | 2001-12-19 | |
| US60/341,978 | 2001-12-19 | ||
| PCT/US2002/041200 WO2003054234A1 (en) | 2001-12-19 | 2002-12-19 | Pcr based high throughput polypeptide screening |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2005512578A true JP2005512578A (ja) | 2005-05-12 |
| JP2005512578A6 JP2005512578A6 (ja) | 2005-08-04 |
| JP2005512578A5 JP2005512578A5 (https=) | 2005-12-22 |
Family
ID=23339812
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2003554934A Pending JP2005512578A (ja) | 2001-12-19 | 2002-12-19 | Ｐｃｒをベースとする高効率のポリペプチドスクリーニング |
Country Status (4)
| Country | Link |
|---|---|
| US (1) | US20030162209A1 (https=) |
| EP (1) | EP1461462A4 (https=) |
| JP (1) | JP2005512578A (https=) |
| WO (1) | WO2003054234A1 (https=) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024248090A1 (ja) * | 2023-05-31 | 2024-12-05 | モデルナ・エンザイマティクス株式会社 | 無細胞系でのｄｎａ種の選択方法及び単離方法 |
| JP7677978B2 (ja) | 2019-12-30 | 2025-05-15 | ナンジン、ジェンスクリプト、バイオテック、カンパニー、リミテッド | 遺伝子変異ライブラリーを構築する方法 |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2513227C (en) * | 2003-01-16 | 2012-09-11 | Genencor International Inc. | Methods for site-directed mutagenesis and targeted randomization |
| EP3351919A1 (en) | 2003-01-24 | 2018-07-25 | University of Utah | Methods of predicting mortality risk by determining telomere length |
| US20100159564A1 (en) * | 2007-11-30 | 2010-06-24 | Dwulet Francis E | Protease resistant recombinant bacterial collagenases |
| CN102439171B (zh) | 2008-12-22 | 2017-07-14 | 犹他大学研究基金会 | 单色多重定量pcr |
| CA2912216A1 (en) | 2013-05-22 | 2014-11-27 | Telomere Diagnostics, Inc. | Measures of short telomere abundance |
| WO2016108954A1 (en) | 2014-12-30 | 2016-07-07 | Telomere Diagnostics, Inc. | Multiplex quantitative pcr |
Family Cites Families (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATE147787T1 (de) * | 1989-07-31 | 1997-02-15 | Inst Of Protein Research Russi | Verfahren zur herstellung von polypeptiden in einem zellfreien system |
| AU638762B2 (en) * | 1989-10-05 | 1993-07-08 | Optein Inc | Cell-free synthesis and isolation of novel genes and polypeptides |
| US5545552A (en) * | 1990-12-03 | 1996-08-13 | Stratagene | Purified thermostable pyrococcus furiosus DNA polymerase I |
| US5866395A (en) * | 1990-12-03 | 1999-02-02 | Stratagene | Purified thermostable pyrococcus furiosus DNA polymerase I |
| WO1993003172A1 (en) * | 1991-08-01 | 1993-02-18 | University Research Corporation | Systematic polypeptide evolution by reverse translation |
| JP3525954B2 (ja) * | 1994-12-27 | 2004-05-10 | 本田技研工業株式会社 | 能動型エンジンマウント装置 |
| US5795961A (en) * | 1995-02-14 | 1998-08-18 | Ludwig Institute For Cancer Research | Recombinant human anti-Lewis b antibodies |
| US5789166A (en) * | 1995-12-08 | 1998-08-04 | Stratagene | Circular site-directed mutagenesis |
| US5848683A (en) * | 1996-07-30 | 1998-12-15 | Seymour; Timothy Harrison | Bucket conveyor chain |
| AU6024598A (en) * | 1997-01-10 | 1998-08-03 | Pioneer Hi-Bred International, Inc. | Hybridization-based genetic amplification and analysis |
| GB9701425D0 (en) * | 1997-01-24 | 1997-03-12 | Bioinvent Int Ab | A method for in vitro molecular evolution of protein function |
| US6183997B1 (en) * | 1997-03-21 | 2001-02-06 | Stratagene | Polymerase enhancing factor (PEF) extracts PEF protein complexes isolated PEF proteins and methods for purifying and identifying same |
| US5923419A (en) * | 1997-06-16 | 1999-07-13 | Insight Control Systems International | System and method for optical inspection of recessed surfaces |
| US6287765B1 (en) * | 1998-05-20 | 2001-09-11 | Molecular Machines, Inc. | Methods for detecting and identifying single molecules |
| US6168931B1 (en) * | 1999-03-17 | 2001-01-02 | The Board Of Trustees Of The Leland Stanford Junior University | Enhanced in vitro synthesis of biological macromolecules using a novel ATP regeneration system |
| US6280977B1 (en) * | 1999-03-24 | 2001-08-28 | Gene Therapy Systems | Method for generating transcriptionally active DNA fragments |
| CA2364572A1 (en) * | 1999-04-12 | 2000-10-19 | Genentech, Inc. | Pcr-based cloning method |
| AU4701200A (en) * | 1999-05-07 | 2000-11-21 | Quantum Dot Corporation | A method of detecting an analyte using semiconductor nanocrystals |
-
2002
- 2002-12-19 US US10/327,292 patent/US20030162209A1/en not_active Abandoned
- 2002-12-19 JP JP2003554934A patent/JP2005512578A/ja active Pending
- 2002-12-19 WO PCT/US2002/041200 patent/WO2003054234A1/en not_active Ceased
- 2002-12-19 EP EP02805672A patent/EP1461462A4/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP7677978B2 (ja) | 2019-12-30 | 2025-05-15 | ナンジン、ジェンスクリプト、バイオテック、カンパニー、リミテッド | 遺伝子変異ライブラリーを構築する方法 |
| WO2024248090A1 (ja) * | 2023-05-31 | 2024-12-05 | モデルナ・エンザイマティクス株式会社 | 無細胞系でのｄｎａ種の選択方法及び単離方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| EP1461462A1 (en) | 2004-09-29 |
| EP1461462A4 (en) | 2006-01-11 |
| WO2003054234A1 (en) | 2003-07-03 |
| US20030162209A1 (en) | 2003-08-28 |
| WO2003054234A9 (en) | 2004-02-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lian et al. | A simple method for developing microsatellite markers using amplified fragments of inter-simple sequence repeat (ISSR) | |
| CN108588050B (zh) | Dna聚合酶以及核酸检测方法和试剂盒 | |
| US10385387B2 (en) | Methods for selectively amplifying and tagging nucleic acids | |
| US20080182296A1 (en) | Pcr-directed gene synthesis from large number of overlapping oligodeoxyribonucleotides | |
| US20060127920A1 (en) | Polynucleotide synthesis | |
| Georgescu et al. | Saturation mutagenesis | |
| TW201321518A (zh) | 微量核酸樣本的庫製備方法及其應用 | |
| JP2017526725A (ja) | 核酸一本鎖環状ライブラリを構築するための方法および試薬 | |
| CN103764849A (zh) | 降低基因组复杂度和多态性检测的方法 | |
| US20210155909A1 (en) | Variant DNA Polymerases Having Improved Properties and Method for Improved Isothermal Amplification of a Target DNA | |
| EP3359669A1 (en) | Devices and methods for producing nucleic acids and proteins | |
| JP6074036B2 (ja) | 拡大された基質範囲を有する新規のｄｎａポリメラーゼ | |
| JP2005512578A (ja) | Ｐｃｒをベースとする高効率のポリペプチドスクリーニング | |
| JP2005512578A6 (ja) | Ｐｃｒをベースとする高効率のポリペプチドスクリーニング | |
| EP1754716B1 (en) | Extreme thermophile single-stranded DNA binding mutant protein, and nucleic acid isothermal amplification method of use thereof | |
| US20230063705A1 (en) | Methods and kits for amplification and detection of nucleic acids | |
| CN116515792B (zh) | Mmlv逆转录酶突变体及其应用 | |
| Miller et al. | A novel, single-tube enzymatic fragmentation and library construction method enables fast turnaround times and improved data quality for microbial whole-genome sequencing | |
| EP2261332A2 (en) | Libraries of recombinant chimeric proteins | |
| US20250051737A1 (en) | Dna polymerase mutant and use thereof | |
| US20210147829A1 (en) | Variant nucleotide library | |
| CN120966792B (zh) | Mmlv逆转录酶突变体、rna逆转录方法及其应用 | |
| US20230340449A1 (en) | Thermostable ligase with reduced sequence bias | |
| EP4039823A1 (en) | Method for micro-volume, one-pot, high-throughput protein production | |
| WO2026071892A1 (en) | N-terminal fragment of dna polymerase, dna polymerase, method for producing the same, and use thereof for isothermal dna amplification at low temperatures |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20050105 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20070522 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20070816 |
|
| A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20070823 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20080205 |