DK173259B1 - Humant papillomavirus type 41-DNA (HPV 41-DNA), diagnostikum indeholdende DNA'et, anvendelse af DNA'et samt fremgangsmåde t - Google Patents
Humant papillomavirus type 41-DNA (HPV 41-DNA), diagnostikum indeholdende DNA'et, anvendelse af DNA'et samt fremgangsmåde t Download PDFInfo
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- DK173259B1 DK173259B1 DK198803834A DK383488A DK173259B1 DK 173259 B1 DK173259 B1 DK 173259B1 DK 198803834 A DK198803834 A DK 198803834A DK 383488 A DK383488 A DK 383488A DK 173259 B1 DK173259 B1 DK 173259B1
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- 238000000034 method Methods 0.000 title claims description 6
- 241000701825 Human papillomavirus type 41 Species 0.000 title abstract description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 5
- 238000001574 biopsy Methods 0.000 claims description 4
- 208000015181 infectious disease Diseases 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 238000010367 cloning Methods 0.000 abstract description 4
- 238000002955 isolation Methods 0.000 abstract description 3
- 206010029098 Neoplasm skin Diseases 0.000 abstract description 2
- 208000000453 Skin Neoplasms Diseases 0.000 abstract description 2
- 238000012512 characterization method Methods 0.000 abstract description 2
- 238000013399 early diagnosis Methods 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 29
- 241000701806 Human papillomavirus Species 0.000 description 28
- 238000003776 cleavage reaction Methods 0.000 description 4
- 230000007017 scission Effects 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 2
- 208000022361 Human papillomavirus infectious disease Diseases 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 208000000260 Warts Diseases 0.000 description 2
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- 229960005542 ethidium bromide Drugs 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 230000008696 hypoxemic pulmonary vasoconstriction Effects 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 201000010153 skin papilloma Diseases 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- 206010059313 Anogenital warts Diseases 0.000 description 1
- 201000009030 Carcinoma 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
- 241000588724 Escherichia coli Species 0.000 description 1
- 208000034179 Neoplasms, Glandular and Epithelial Diseases 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 241001631646 Papillomaviridae Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
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- 239000011543 agarose gel Substances 0.000 description 1
- 102000005936 beta-Galactosidase Human genes 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- AIYUHDOJVYHVIT-UHFFFAOYSA-M caesium chloride Chemical compound [Cl-].[Cs+] AIYUHDOJVYHVIT-UHFFFAOYSA-M 0.000 description 1
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- 108091092356 cellular DNA Proteins 0.000 description 1
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- 238000003745 diagnosis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000004392 genitalia Anatomy 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
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- 230000008018 melting Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 201000008261 skin carcinoma Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- PIEPQKCYPFFYMG-UHFFFAOYSA-N tris acetate Chemical compound CC(O)=O.OCC(N)(CO)CO PIEPQKCYPFFYMG-UHFFFAOYSA-N 0.000 description 1
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- 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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P35/00—Antineoplastic agents
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- 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
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
- C12Q1/701—Specific hybridization probes
- C12Q1/708—Specific hybridization probes for papilloma
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- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K39/00—Medicinal preparations containing antigens or antibodies
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- C12N2710/00011—Details
- C12N2710/20011—Papillomaviridae
- C12N2710/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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Description
DK 173259 B1 i
Den foreliggende opfindelse angår humant papillomavirus type 41-DNA, et diagnostikum indeholdende DNA'et, anvendelsen af DNA'et samt en fremgangsmåde til diagnosticering af HPV 41-infektioner.
t 5 Humane papillomaviruser (HPV) danner en gruppe på ca. 40 forskellige typer (H. zur Hausen og A. Schneider, (1987), The Papillomaviruses, P.M. Howley og N.P. Salzmann, ed.). HPV er fundet i sammenhæng med godartede (vorter, kon-dylomer i genitalområdet) og ondartede (carcinomer i hud og 10 skede) epitheliale neoplasmer. Papillomaviruser kan ikke formeres i kultur. Anvendelsen af humant papillomavirus type 41-DNA (HPV 41-DNA) som diagnostikum samt udvinding af ekspressionsprodukter forudsætter således genteknologiske fremgangsmåder.
15 Det har været formålet med opfindelsen at tilveje bringe sådanne genteknologiske fremgangsmåder.
Til grund for opfindelsen ligger isolering for første gang af HPV 41, en delvis karakterisering af dets genom samt kloning i pUC 19. Herved åbnes mulighed for tidlig 20 diagnose af hudtumorer, der er knyttet til HPV 41.
Opfindelsen angår således HPV41-DNA, som er ejendommeligt ved, at det er en klon med deponeringsnummeret DSM 4175 P eller DSM 4174 P.
Opfindelsen angår også et diagnostikum, som er ejen-25 dommeligt ved, at det indeholder DNA ifølge opfindelsen eller dele deraf.
Opfindelsen angår desuden anvendelsen af DNA ifølge opfindelsen til genteknologisk ekspression af proteiner.
Opfindelsen angår endvidere en fremgangsmåde til 30 diagnosticering af HPV 41-infektioner, hvilken fremgangsmåde er ejendommelig ved, at biopsimateriale eller afskrabet materiale bringes i kontakt med et diagnostikum ifølge opfindelsen, eller at RNA eller DNA, som skal undersøges, hybridiseres med DNA ifølge opfindelsen.
35 Yderligere udførelsesformer for opfindelsen er be skrevet nærmere i det følgende.
2 DK 173259 B1
Kloningen af HPV 41 har muliggjort sammenligning med 40 andre HPV. HPV 7, 8, 10, 17, 27, 29, 30 og 33 er fjernt beslægtede, idet der er påvist kolinearitet af HPV 41 med HPV 8 (fig. 2) og er tilvejebragt fysiske genomkort for 5 restriktionsenzymspaltninger (fig. 1).
Der er dermed åbnet mulighed for at undersøge neopla-sier, især carcinomer i huden, for forekomst af HPV 41.
Eksempler 10 1. Isolering af episomalt HPV 41-DNA.
Biopsier fra vortevæv fra tre forskellige hudarealer hos en 15-årig pige dybfryses umiddelbart efter tilvejebringelsen ved -70eC og opbevares. Deraf isoleres højmolekylært DNA som beskrevet af Gissmann et al. (1982), Int. J. Cancer 15 29, 143-146. Ringformigt lukket dobbeltstrenget DNA udvindes fra det nævnte cellulære DNA ifølge Radioff et al. (1967)
Proc. Nat. Acad. Sci. 57, 1514-1521, idet ca. 10 /*g DNA
centrifugeres i en 50 Ti-rotor (Beckmann Corp.) ved 45.000 o/m i 48 timer i cæsiumchlorid (vægtfylde 1,56 g/ml) med til-20 sætning af 60 0 /ig/ml ethidiumbromid, hvorefter der opsamles fraktioner med en vægtfylde på 1,59-1,60. Alternativt kan episomalt HPV 41-DNA af ikke-spaltet DNA fra det samme biop-simateriale påvises i agarosegeler farvet med ethidiumbromid (1% agarose (Seakem ME) i 40 mM Tris-acetat, 2 mM EDTA (pH-25 værdi 7,8)) og isoleres derfra.
2. Kloning af HPV 41 i plasmid pUC 19.
Som kloningsvektor vælges det kendte plasmid pUC 19 (Yanish-Perron et al. (1985), Gene 33.» 113-119). Cirkulært 30 lukket dobbeltstrenget DNA klones i pUC 19 efter spaltning med BamHI (Maniatis et al. (1982), Molecular Cloning: A
Laboratory Manual; Cold Spring Harbor Laboratory Press, New York) . Rekombinante kloner identificeres ved en β-galacto-sidase-test (Messing et al. (1977), Proc. Nat. Acad. Sci.
3 5 USA 74., 364 2-3646) , og indligerede DNA-stykker analyseres efter DNA-hurtigekstraktion (H.L. Birnboim og Doly. (1979) , 3 DK 173259 B1
Nuel. Acids Res. 7, 1513-1523).
Der fås to rekombinanter, hvoraf den ene, K 10, indeholder en indsats på 6,6 kb, og den anden, K 6, indeholder en indsats på 0,98 kb. Homologien af disse klonede sekvenser , 5 er påvist ved hybridisering med episomalt DNA fra biopsimate- riale.
3. Fysiske genomkort af HPV 41.
HVP 41-DNA fraskilt fra vektoren ved BamHI-spaltning 10 nedbrydes med restriktionsendonukleaserne Accl, Bglll, Xbal,
Smal, EcoRI, PstI og Hindi, og de tilsvarende fysiske genomkort tilvejebringes ved almindeligt kendte metoder. Resultaterne er sammenfattet i fig. l, hvor det singulære Kpnl--spaltningssted tjener til at linearisere HPV 41-molekylet.
15 4. Sammenligning med andre HPV.
DNA af HPV 41-genomet sammenlignes med DNA’er af 40 tilgængelige HPV-typer ved hjælp af DNA/DNA-hybridisering under forskellige stringens (E.M. Southern (1975), J. Mol.
20 Biol. j?8, 503-517). Under betingelser med høj stringens (smeltetemperatur Tm = -20°C) hybridiseres HPV 41-K 10-DNA delvis med HPV 29-DNA. DNA af K 6-klonen hybridiserer kun ved lav stringens (Tm = -40°C) med HPV 3- og 13-DNA.
Kolineariteten med HPV 8 på grundlag af hybridise-25 ringsforsøg er vist i fig. 2.
Med kendskab til HPV 8-DNA-sekvensen (Fuchs et al.
(1986), J. Virol. 58, 626-634) kan den åbne læseramme af HPV 41 afledes, og dermed bliver det muligt at få HPV 41--proteinerne ved almindeligt kendte metoder til subkloning 30 af klonerne K 10 og K 6 med efterfølgende ekspression i prokaryotiske eller eukaryotiske ekspressionssystemer.
Plasmiderne HPV 41, klon K 6 og K 10 er deponeret (i E. coli) den 3. juli 1987 hos Deutsche Sammlung fur Mikroorganismen under numrene DSM 4174P (K 6) og DSM 4175P (K 10) .
Claims (4)
1. HPV 41-DNA, kendetegnet ved, at det er en klon med deponeringsnummeret DSM 4175 P eller DSM 4174 P.
2. Diagnostikum, kendetegnet ved, at det indholder DNA ifølge krav 1 eller dele deraf.
3. Anvendelse af DNA ifølge krav l til genteknologisk ekspression af proteiner.
4. Fremgangsmåde til diagnosticering af HPV 41-infek-10 tioner, kendetegnet ved, at biopsimateriale eller afskrabet materiale bringes i kontakt med et diagnostikum ifølge krav 2, eller at RNA eller DNA, som skal undersøges, hybridiseres med DNA ifølge krav l.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE3722968 | 1987-07-11 | ||
| DE19873722968 DE3722968A1 (de) | 1987-07-11 | 1987-07-11 | Humaner papillomvirus typ 41, seine dna und die dafuer kodierenden proteine |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| DK383488D0 DK383488D0 (da) | 1988-07-08 |
| DK383488A DK383488A (da) | 1989-01-12 |
| DK173259B1 true DK173259B1 (da) | 2000-05-29 |
Family
ID=6331386
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK198803834A DK173259B1 (da) | 1987-07-11 | 1988-07-08 | Humant papillomavirus type 41-DNA (HPV 41-DNA), diagnostikum indeholdende DNA'et, anvendelse af DNA'et samt fremgangsmåde t |
Country Status (12)
| Country | Link |
|---|---|
| US (2) | US5142032A (da) |
| EP (1) | EP0301289B1 (da) |
| JP (1) | JP2746380B2 (da) |
| KR (1) | KR970005047B1 (da) |
| AT (1) | ATE111158T1 (da) |
| AU (1) | AU617330B2 (da) |
| CA (1) | CA1340322C (da) |
| DE (2) | DE3722968A1 (da) |
| DK (1) | DK173259B1 (da) |
| ES (1) | ES2060625T3 (da) |
| FI (1) | FI100538B (da) |
| PT (1) | PT87940B (da) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK1314431T3 (da) * | 1993-04-30 | 2008-11-10 | Wellstat Biologics Corp | Oprensede sammensætninger af Newcastle disease virus |
| DE19526386C1 (de) | 1995-07-19 | 1997-01-02 | Deutsches Krebsforsch | Papillomviren, Mittel zu deren Nachweis sowie zur Therapie von durch sie verursachten Erkrankungen |
| JP2002541822A (ja) * | 1999-04-14 | 2002-12-10 | エム ユー エス シー ファンデーション フォー リサーチ ディベロップメント | 組織特異的および病原体特異的毒性物質ならびにリボザイム |
| US20030059806A1 (en) * | 2001-01-05 | 2003-03-27 | Science & Technology Corporation @ Unm | Probes for the detection of human papillomavirus |
| CA2491034A1 (en) * | 2002-06-26 | 2004-01-08 | The Penn State Research Foundation | Methods and materials for treating human papillomavirus infections |
| WO2011149897A1 (en) | 2010-05-25 | 2011-12-01 | Qiagen Gaithersburg, Inc. | Fast results hybrid capture assay and associated strategically-truncated probes |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1986005816A1 (en) * | 1985-04-04 | 1986-10-09 | Georgetown University | Type-specific papillomavirus dna sequences and peptides |
| DE3625257A1 (de) * | 1986-07-23 | 1988-02-04 | Behringwerke Ag | Expressionsprodukte der menschlichen papillomviren typ 16 und 18, fuer diese proteine spezifische antikoerper und diese antikoerper bzw. entsprechende dna enthaltende diagnostika |
| US4849331A (en) * | 1987-06-09 | 1989-07-18 | Life Technologies, Inc. | Human papillomavirus 44 nucleic acid hybridization probes and methods for employing the same |
| US4849334A (en) * | 1987-06-09 | 1989-07-18 | Life Technologies, Inc. | Human papillomavirus 43 nucleic acid hybridization probes and methods for employing the same |
| DE3722967A1 (de) * | 1987-07-11 | 1989-01-19 | Behringwerke Ag | Monoklonale antikoerper gegen e7-protein des humanen papillomvirus typ 16, verfahren zu ihrer herstellung sowie ihre verwendung |
| DE3826793A1 (de) * | 1988-08-06 | 1990-03-08 | Behringwerke Ag | Humaner papillomvirus typ 57, seine dna und die davon kodierten proteine |
-
1987
- 1987-07-11 DE DE19873722968 patent/DE3722968A1/de not_active Withdrawn
-
1988
- 1988-07-07 DE DE3851391T patent/DE3851391D1/de not_active Expired - Fee Related
- 1988-07-07 ES ES88110851T patent/ES2060625T3/es not_active Expired - Lifetime
- 1988-07-07 FI FI883255A patent/FI100538B/fi not_active IP Right Cessation
- 1988-07-07 EP EP88110851A patent/EP0301289B1/de not_active Expired - Lifetime
- 1988-07-07 AT AT88110851T patent/ATE111158T1/de not_active IP Right Cessation
- 1988-07-08 PT PT87940A patent/PT87940B/pt not_active IP Right Cessation
- 1988-07-08 CA CA000571554A patent/CA1340322C/en not_active Expired - Fee Related
- 1988-07-08 US US07/216,913 patent/US5142032A/en not_active Expired - Lifetime
- 1988-07-08 AU AU18878/88A patent/AU617330B2/en not_active Ceased
- 1988-07-08 DK DK198803834A patent/DK173259B1/da not_active IP Right Cessation
- 1988-07-11 KR KR1019880008579A patent/KR970005047B1/ko not_active IP Right Cessation
- 1988-07-11 JP JP63172512A patent/JP2746380B2/ja not_active Expired - Fee Related
-
1993
- 1993-11-23 US US08/156,936 patent/US5382509A/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6455182A (en) | 1989-03-02 |
| KR970005047B1 (ko) | 1997-04-11 |
| EP0301289B1 (de) | 1994-09-07 |
| US5382509A (en) | 1995-01-17 |
| PT87940B (pt) | 1995-03-01 |
| CA1340322C (en) | 1999-01-19 |
| FI883255A (fi) | 1989-01-12 |
| US5142032A (en) | 1992-08-25 |
| AU1887888A (en) | 1989-04-20 |
| DE3851391D1 (de) | 1994-10-13 |
| FI883255A0 (fi) | 1988-07-07 |
| EP0301289A1 (de) | 1989-02-01 |
| DK383488A (da) | 1989-01-12 |
| JP2746380B2 (ja) | 1998-05-06 |
| DE3722968A1 (de) | 1989-01-19 |
| ES2060625T3 (es) | 1994-12-01 |
| ATE111158T1 (de) | 1994-09-15 |
| FI100538B (fi) | 1997-12-31 |
| AU617330B2 (en) | 1991-11-28 |
| DK383488D0 (da) | 1988-07-08 |
| PT87940A (pt) | 1989-06-30 |
| KR890002402A (ko) | 1989-04-10 |
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