JPH0129557B2 - - Google Patents
Info
- Publication number
- JPH0129557B2 JPH0129557B2 JP55033931A JP3393180A JPH0129557B2 JP H0129557 B2 JPH0129557 B2 JP H0129557B2 JP 55033931 A JP55033931 A JP 55033931A JP 3393180 A JP3393180 A JP 3393180A JP H0129557 B2 JPH0129557 B2 JP H0129557B2
- Authority
- JP
- Japan
- Prior art keywords
- dna
- mrna
- interferon
- plasmid
- coli
- 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
Links
- 108020004999 messenger RNA Proteins 0.000 claims description 71
- 239000013612 plasmid Substances 0.000 claims description 54
- 241000588724 Escherichia coli Species 0.000 claims description 23
- 102000003996 Interferon-beta Human genes 0.000 claims description 21
- 108090000467 Interferon-beta Proteins 0.000 claims description 21
- 108090000623 proteins and genes Proteins 0.000 claims description 16
- 108020004414 DNA Proteins 0.000 description 80
- 102000053602 DNA Human genes 0.000 description 79
- 108010050904 Interferons Proteins 0.000 description 37
- 102000014150 Interferons Human genes 0.000 description 35
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 34
- 229940079322 interferon Drugs 0.000 description 34
- 239000000243 solution Substances 0.000 description 21
- 238000000034 method Methods 0.000 description 20
- 239000011780 sodium chloride Substances 0.000 description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 15
- 108091008146 restriction endonucleases Proteins 0.000 description 15
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 14
- 229920002477 rna polymer Polymers 0.000 description 14
- 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 12
- 210000002950 fibroblast Anatomy 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 11
- 230000000295 complement effect Effects 0.000 description 11
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 10
- 239000000872 buffer Substances 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- -1 30mM) Chemical compound 0.000 description 8
- 102100034343 Integrase Human genes 0.000 description 8
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 8
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 6
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 6
- 229960000723 ampicillin Drugs 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 description 5
- 102100029764 DNA-directed DNA/RNA polymerase mu Human genes 0.000 description 5
- 239000000020 Nitrocellulose Substances 0.000 description 5
- 230000000840 anti-viral effect Effects 0.000 description 5
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethyl mercaptane Natural products CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 5
- 229920001220 nitrocellulos Polymers 0.000 description 5
- 210000000287 oocyte Anatomy 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 5
- 108020004682 Single-Stranded DNA Proteins 0.000 description 4
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 4
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 108060002716 Exonuclease Proteins 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- 108020004511 Recombinant DNA Proteins 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 241000269370 Xenopus <genus> Species 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 108091092330 cytoplasmic RNA Proteins 0.000 description 3
- RGWHQCVHVJXOKC-SHYZEUOFSA-N dCTP Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](CO[P@](O)(=O)O[P@](O)(=O)OP(O)(O)=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-N 0.000 description 3
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 3
- 102000013165 exonuclease Human genes 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 201000008968 osteosarcoma Diseases 0.000 description 3
- HJRIWDYVYNNCFY-UHFFFAOYSA-M potassium;dimethylarsinate Chemical compound [K+].C[As](C)([O-])=O HJRIWDYVYNNCFY-UHFFFAOYSA-M 0.000 description 3
- 239000012266 salt solution Substances 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000701959 Escherichia virus Lambda Species 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 108091005904 Hemoglobin subunit beta Proteins 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 241000711408 Murine respirovirus Species 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical group O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 238000000376 autoradiography Methods 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 230000016396 cytokine production Effects 0.000 description 2
- 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 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 2
- 230000002285 radioactive effect Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 102100027211 Albumin Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 241000713838 Avian myeloblastosis virus Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108090000204 Dipeptidase 1 Proteins 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 241001235200 Haemophilus influenzae Rd KW20 Species 0.000 description 1
- 102100021519 Hemoglobin subunit beta Human genes 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 101710149086 Nuclease S1 Proteins 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 Teflon® Polymers 0.000 description 1
- 241000269368 Xenopus laevis Species 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000000211 autoradiogram Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 102000006635 beta-lactamase Human genes 0.000 description 1
- 239000007978 cacodylate buffer Substances 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 210000003953 foreskin Anatomy 0.000 description 1
- 102000018146 globin Human genes 0.000 description 1
- 108060003196 globin Proteins 0.000 description 1
- 238000003505 heat denaturation Methods 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/555—Interferons [IFN]
- C07K14/565—IFN-beta
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Priority Applications (14)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3393180A JPS56131598A (en) | 1980-03-19 | 1980-03-19 | Novel recombinant plasmid having gene of human fibroblastic interferon messenger rna |
AU63762/80A AU538665B2 (en) | 1979-10-30 | 1980-10-28 | Human interferon dna |
IL61366A IL61366A (en) | 1979-10-30 | 1980-10-29 | Dna with interferon activity,recombinant plasmids containing it,microorganisms prepared therewith and methods for the preparation thereof |
DK458480A DK458480A (da) | 1979-10-30 | 1980-10-29 | Dna, recombinant plasmid og mikroorganisme indeholdende dette samt fremgangsmaader til fremstilling deraf |
NO803230A NO158950C (no) | 1979-10-30 | 1980-10-29 | Dna-sekvens som koder for et polypeptid med human fibroblast-interferonaktivitet, fremgangsmaate for dens fremstilling, samt fremgangsmaate for fremstilling av et plasmid. |
AT80106685T ATE18072T1 (de) | 1979-10-30 | 1980-10-30 | Doppelstraengige dns, welche fuer ein polypeptid mit der aktivitaet von menschlichem fibroblasten- interferon codiert, clonierte dns, rekombinantes plasmid, welches die dns enthaelt, und mikroorganismus, welcher das rekombinante plasmid enthaelt. |
DE8080106685T DE3071435D1 (en) | 1979-10-30 | 1980-10-30 | Double-stranded dna which codes for a polypeptide with human fibroblast interferon activity, cloned dna, recombinant plasmid containing the dna and microorganism containing the recombinant plasmid |
EP80106685A EP0028033B1 (de) | 1979-10-30 | 1980-10-30 | Doppelsträngige DNS, welche für ein Polypeptid mit der Aktivität von menschlichem Fibroblasten-Interferon codiert, clonierte DNS, rekombinantes Plasmid, welches die DNS enthält, und Mikroorganismus, welcher das rekombinante Plasmid enthält |
ES496426A ES8203102A1 (es) | 1979-10-30 | 1980-10-30 | Un procedimiento para la preparacion de un adn que codifica un polipeptido con actividad de interferon. |
CA000363628A CA1341563C (en) | 1979-10-30 | 1980-10-30 | Dna coding for human fibroblast interferon polypeptide and recombinant plasmid |
ES507596A ES8303530A1 (es) | 1979-10-30 | 1981-11-30 | Un procedimiento para la preparacion de un microorganismo que contiene un plasmido recombinante de un adn |
ES507595A ES8303529A1 (es) | 1979-10-30 | 1981-11-30 | Un procedimiento para la produccion de un plasmido recombi- nante. |
US08/400,179 US5514567A (en) | 1979-01-30 | 1995-03-06 | DNA and recombinant plasmid |
US08/463,757 US9376478B1 (en) | 1979-10-30 | 1995-06-05 | DNA and recombinant plasmid |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3393180A JPS56131598A (en) | 1980-03-19 | 1980-03-19 | Novel recombinant plasmid having gene of human fibroblastic interferon messenger rna |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS56131598A JPS56131598A (en) | 1981-10-15 |
JPH0129557B2 true JPH0129557B2 (de) | 1989-06-12 |
Family
ID=12400254
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP3393180A Granted JPS56131598A (en) | 1979-01-30 | 1980-03-19 | Novel recombinant plasmid having gene of human fibroblastic interferon messenger rna |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS56131598A (de) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3177213D1 (de) | 1980-04-03 | 1990-10-18 | Biogen Inc | Dns-sequenzen, rekombinante dns-molekuele und verfahren zur herstellung von dem menschlichen fibroblast-interferon. |
IE54592B1 (en) * | 1982-03-08 | 1989-12-06 | Genentech Inc | Anumal interferons, processes involved in their production, compositions containing them, dna sequences coding therefor and espression vehicles containing such sequences and cells transformed thereby |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5685296A (en) * | 1979-11-21 | 1981-07-11 | Yeda Res & Dev | Isolation of mrna convertible to interferon and corresponding dna and production of interferonpolypeptide and high purity interferon |
-
1980
- 1980-03-19 JP JP3393180A patent/JPS56131598A/ja active Granted
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5685296A (en) * | 1979-11-21 | 1981-07-11 | Yeda Res & Dev | Isolation of mrna convertible to interferon and corresponding dna and production of interferonpolypeptide and high purity interferon |
Also Published As
Publication number | Publication date |
---|---|
JPS56131598A (en) | 1981-10-15 |
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