DE3320339C2 - - Google Patents
Info
- Publication number
- DE3320339C2 DE3320339C2 DE3320339A DE3320339A DE3320339C2 DE 3320339 C2 DE3320339 C2 DE 3320339C2 DE 3320339 A DE3320339 A DE 3320339A DE 3320339 A DE3320339 A DE 3320339A DE 3320339 C2 DE3320339 C2 DE 3320339C2
- Authority
- DE
- Germany
- Prior art keywords
- dna
- binding protein
- gene
- phosphate binding
- dna fragment
- 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
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- 239000012634 fragment Substances 0.000 claims description 73
- 108090000623 proteins and genes Proteins 0.000 claims description 63
- 230000009466 transformation Effects 0.000 claims description 60
- 108010058514 Phosphate-Binding Proteins Proteins 0.000 claims description 47
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- 102000006335 Phosphate-Binding Proteins Human genes 0.000 claims description 46
- 239000013598 vector Substances 0.000 claims description 45
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- 238000005215 recombination Methods 0.000 claims description 38
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- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
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- 239000003184 complementary RNA Substances 0.000 description 1
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- 229960000258 corticotropin Drugs 0.000 description 1
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- 230000018109 developmental process Effects 0.000 description 1
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- 239000012153 distilled water Substances 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
- 229940079593 drug Drugs 0.000 description 1
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- 239000011874 heated mixture Substances 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
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- 238000003384 imaging method Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 108010031622 lead-binding proteins Proteins 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 229960004452 methionine Drugs 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000002636 mycotoxin Substances 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000003399 opiate peptide Substances 0.000 description 1
- 229940092253 ovalbumin Drugs 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000009938 salting Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000003998 snake venom 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
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 1
- 229960000553 somatostatin Drugs 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- WPLOVIFNBMNBPD-ATHMIXSHSA-N subtilin Chemical compound CC1SCC(NC2=O)C(=O)NC(CC(N)=O)C(=O)NC(C(=O)NC(CCCCN)C(=O)NC(C(C)CC)C(=O)NC(=C)C(=O)NC(CCCCN)C(O)=O)CSC(C)C2NC(=O)C(CC(C)C)NC(=O)C1NC(=O)C(CCC(N)=O)NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C1NC(=O)C(=C/C)/NC(=O)C(CCC(N)=O)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)CNC(=O)C(NC(=O)C(NC(=O)C2NC(=O)CNC(=O)C3CCCN3C(=O)C(NC(=O)C3NC(=O)C(CC(C)C)NC(=O)C(=C)NC(=O)C(CCC(O)=O)NC(=O)C(NC(=O)C(CCCCN)NC(=O)C(N)CC=4C5=CC=CC=C5NC=4)CSC3)C(C)SC2)C(C)C)C(C)SC1)CC1=CC=CC=C1 WPLOVIFNBMNBPD-ATHMIXSHSA-N 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
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- 239000012137 tryptone Substances 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 229960005356 urokinase Drugs 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-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
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
- C12N9/2471—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01023—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/034—Fusion polypeptide containing a localisation/targetting motif containing a motif for targeting to the periplasmic space of Gram negative bacteria as a soluble protein, i.e. signal sequence should be cleaved
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/61—Fusion polypeptide containing an enzyme fusion for detection (lacZ, luciferase)
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/822—Microorganisms using bacteria or actinomycetales
- Y10S435/848—Escherichia
- Y10S435/849—Escherichia coli
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Description
- 1) Gewinnung einer chromosomalen DNS (DNA) mit einer Genkodierung für ein Phosphatbindungsprotein aus Bakterien der Familie Enterobacteriaceae;
- 2) Aufspaltung der chromosomalen DNS mit Hilfe eines Re striktionsenzyms in DNS-Fragmente;
- 3) Ligieren der DNS-Fragmente an ein Replikon aus der Gruppe der Plasmide;
- 4) Transformierung der Zellen eines Bakteriums aus der Familie Enterobacteriaceae mit Hilfe des Replikons, an welches das DNS-Fragment ligiert ist, in Transformationsprodukte, darunter Transformationsprodukte, welche einen Rekombinationsvektor mti dem DNS-Fragment enthalten, das eine Genkodierung für ein Phosphatbindungsprotein mit sich führt;
- 5) Auslesen von Transformationsprodukten, welche den Re kombinationsvektor mit dem eine Genkodierung für ein Phosphatbindungsprotein enthaltenden DNS-Fragment beinhalten, aus den Transformationsprodukten, und
- 6) Isolieren des Rekombinationsvektors mit dem die Genkodierung für ein Phosphatbindungsprotein führenden DNS-Fragment aus den ausgelesenen bzw. selektierten Transformationsprodukten.
EcoRI, HpaI, PstI, BglII, MluI HindIII, AvaI, HincII und SmaI (wobei die angehängten Zahlen bei diesen Symbolen gemäß der Zeichnung zur Kennzeichnung der jeweiligen Erkennungsorte für ein Restriktionsenzym eingesetzt sind.)
Tn3(ApR) (angegeben mit ): Transposon
welches eine Genkodierung für die Ampicillinresistenz enthält;
Rep: Replikationsgen
ori: Replikationsausgangspunkt
Kb: 1000 Nukleotidenpaare
: ausgelöschter Teil
- 1) eine Chromosomen-DNS hergestellt wird, die eine Genkodierung für ein Protein zur Phosphatbindung enthält, und zwar aus einem Bakterium der Familie der Enterobac teriaceen;
- 2) die Chromosomen-DNS mit einem Restriktionsenzym zur Erzeugung von DNS-Fragmenten gespalten wird;
- 3) die DNS-Fragmente an ein aus der Gruppe der Plasmide und Bakteriophagen gewähltes Replikon ligiert werden;
- 4) Zellen eines Bakteriums aus der Familie der Enterobacteriaceen mit dem Replikon, an welches die DNS-Fragmente ligiert sind, zur Bildung von Transformationsprodukten transformiert werden, darunter solchen, die einen Rekombinationsvektor enthalten, unter anderem die DNS-Fragmente, welche eine Genkodierung für ein Phosphat bindungsprotein führen;
- 5) Transformationsprodukte, welche den Rekombinationsvektor, darunter die eine Genkodierung für ein Phosphatbindungsprotein führenden DNS-Fragmente, aus den gesamten Transformationsprodukten ausgewählt werden, und
- 6) der Rekombinationsvektor, einschließlich der die Genkodierung für ein Phosphatbindungsprotein führenden DNS-Fragmente, aus den ausgewählten Transformationsprodukten isoliert wird.
- 1) Die Ausbeute des vorgenannten erwünschten Polypeptides beträgt 1×10⁵-10⁶ Moleküle/Zelle, also mehrere bis 100Male höher als bei Verwendung des üblicherweise verwendeten Vektors.
- 2) Die Genexpression des Expressionsvektors gemäß der vorliegenden Erfindung läßt sich einschränkungslos durch Einstellung der Phosphatkonzentration im Kulturmedium steuern. Damit läßt sich auch die Produktion des gewünschten Polypeptides durch Einstellen der Phosphatkonzentration im Kulturmedium steuern.
- 3) Das gewünschte Polypeptid kann zusammen mit dem Anteil an Phosphatbindungsprotein in das Periplasma des Wirts ausscheiden. Damit läßt sich das gewünschte Polypeptid ohne Schwierigkeiten vom Wirt isolieren.
A: Zusammensetzung des T-Mediums | |
Bacto-Trypton|10 g | |
NaCl | 5 g |
D: Zusammensetzung des Puffers für das EcoRI-Restriktionsenzym (fünffach konzentriert) | |
NaCl|500 mM | |
Tris-HCl | 250 mM (pH 7,4) |
MgSO₄ | 50 mM |
E: Zusammensetzung des Ligasepuffers | |
Tris-HCl | |
66 mM (pH 7,6) | |
MgCl₂ | 6,6 mM |
Dithiothreitol | 10 mM |
ATP | 0,5 mM |
A = Ausbeute an Phosphatbindungsprotein (g/Zelle)
B = Molekulargewicht des Phosphatbindungs proteins (35 000)
C = Avogadro′sche Zahl (6×10²³).
Claims (7)
- 1) Gewinnung einer chromosomalen DNS mit einer Genkodierung für ein Phosphatbindungsprotein aus Bakterien der Familie Enterobacteriaceae gehören;
- 2) Aufspaltung der Chromosomen-DNS mit Hilfe eines Restriktionsenzyms in DNS-Fragmente;
- 3) Ligieren der DNS-Fragmente an ein Replikon aus der Gruppe der Plasmide und Bakteriophagen;
- 4) Transformierung der Zellen eines Bakteriums aus der Familie Enterobacteriaceae mit Hilfe des Replikons, an welches das DNS-Fragment ligiert ist, in Transformationsprodukte, darunter Transformationsprodukte, welche einen Rekombinationsvektor mit dem DNS-Fragment enthalten, das eine Genkodierung für ein Phosphatbindungsprotein mit sich führt;
- 5) Auslesen von Transformationsprodukten, welche den Rekombinationsvektor mit dem eine Genkodierung für ein Phosphatbindungsprotein enthaltenden DNS-Fragment beinhalten, aus den Transformationsprodukten, und
- 6) Isolieren des Rekombinationsvektors mit dem die Genkodierung für ein Phosphatbindungsprotein führenden DNS-Fragment aus den ausgelesenen Transformationspro dukten.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP57096775A JPS592689A (ja) | 1982-06-04 | 1982-06-04 | 強力な遺伝子発現能を有する新規レプリコンの作成法 |
Publications (2)
Publication Number | Publication Date |
---|---|
DE3320339A1 DE3320339A1 (de) | 1984-01-19 |
DE3320339C2 true DE3320339C2 (de) | 1990-08-23 |
Family
ID=14174007
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DE19833320339 Granted DE3320339A1 (de) | 1982-06-04 | 1983-06-04 | Expressionsvektor, verfahren zu seiner herstellung und verwendung des expressionsvektors zur herstellung eines polypeptids |
Country Status (9)
Country | Link |
---|---|
US (1) | US4703005A (de) |
JP (1) | JPS592689A (de) |
BE (1) | BE896957A (de) |
CA (1) | CA1202258A (de) |
DE (1) | DE3320339A1 (de) |
FR (1) | FR2528070B1 (de) |
GB (1) | GB2124232B (de) |
NL (1) | NL189208C (de) |
SE (1) | SE459816B (de) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0699580B2 (ja) * | 1986-05-28 | 1994-12-07 | 花王株式会社 | 多孔性フイルム |
FR2599380A1 (fr) * | 1986-05-29 | 1987-12-04 | Centre Nat Rech Scient | Vecteur d'exportation de proteines chez escherichiacoli, bacteries transformees et procede pour la preparation de proteines |
JPH0699581B2 (ja) * | 1986-06-09 | 1994-12-07 | 花王株式会社 | 多孔性フイルム |
JPH0768394B2 (ja) * | 1987-12-01 | 1995-07-26 | 花王株式会社 | 多孔性フィルム及びその製造方法 |
US5139954A (en) * | 1988-03-29 | 1992-08-18 | E. I. Du Pont De Nemours And Company | DNA promoter fragments from wheat |
NZ228320A (en) | 1988-03-29 | 1991-06-25 | Du Pont | Nucleic acid promoter fragments of the promoter region homologous to the em gene of wheat, dna constructs therefrom and plants thereof |
KR920701453A (ko) * | 1989-03-17 | 1992-08-11 | 미리엄 디. 멕코나헤이 | 유전자발현의 외부조절 |
US5304472A (en) * | 1992-11-20 | 1994-04-19 | Genentech, Inc. | Method of controlling polypeptide production in bacterial cells |
US5965402A (en) * | 1996-02-20 | 1999-10-12 | Smithkline Beecham Corporation | DNA encoding phoH polypeptides |
JP4229341B2 (ja) | 1996-03-23 | 2009-02-25 | 財団法人阪大微生物病研究会 | 破傷風毒素の機能的フラグメント抗原及び破傷風ワクチン |
AU2016253217B2 (en) * | 2015-04-24 | 2021-09-02 | Ajinomoto Co., Inc. | Method for secretory production of protein |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4237224A (en) * | 1974-11-04 | 1980-12-02 | Board Of Trustees Of The Leland Stanford Jr. University | Process for producing biologically functional molecular chimeras |
GB2023612B (en) * | 1978-06-01 | 1982-09-15 | Hopwood D A | Inc of nucleic acid into cellular systems streptomyces plasmids |
US4411994A (en) * | 1978-06-08 | 1983-10-25 | The President And Fellows Of Harvard College | Protein synthesis |
FR2441659A1 (fr) * | 1978-11-14 | 1980-06-13 | Anvar | Nouveaux plasmides hybrides et microorganismes les contenant |
US4273875A (en) * | 1979-03-05 | 1981-06-16 | The Upjohn Company | Plasmid and process of isolating same |
AU542264B2 (en) * | 1979-06-01 | 1985-02-14 | G.D. Searle & Co. | Plasmid vectors |
DE2931999A1 (de) * | 1979-08-03 | 1981-02-26 | Schering Ag | Herstellung und anwendung von neukombinierten plasmiden mit genen fuer alkalische phosphatasen |
NO159863C (no) * | 1980-01-07 | 1989-02-15 | Univ Rochester | Fremgangsm te for fremstilling og seleksjon av en rant bakteriofag som inneholder et genetisk fragment og koder for alfa-amylase, egnet for bruk i heterolog transformering av en bacillus-verts-mikroorganisme. |
US4374927A (en) * | 1981-02-24 | 1983-02-22 | The Board Of Trustees Of The Leland Stanford Jr. University | Extrachromosomal regulation of expression |
-
1982
- 1982-06-04 JP JP57096775A patent/JPS592689A/ja active Granted
-
1983
- 1983-05-25 CA CA000428884A patent/CA1202258A/en not_active Expired
- 1983-05-31 SE SE8303068A patent/SE459816B/sv not_active IP Right Cessation
- 1983-06-03 NL NLAANVRAGE8301986,A patent/NL189208C/xx not_active IP Right Cessation
- 1983-06-03 BE BE0/210930A patent/BE896957A/fr not_active IP Right Cessation
- 1983-06-03 GB GB08315278A patent/GB2124232B/en not_active Expired
- 1983-06-03 FR FR8309292A patent/FR2528070B1/fr not_active Expired
- 1983-06-04 DE DE19833320339 patent/DE3320339A1/de active Granted
- 1983-06-06 US US06/501,559 patent/US4703005A/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
BE896957A (fr) | 1983-12-05 |
JPH0116159B2 (de) | 1989-03-23 |
FR2528070B1 (fr) | 1987-07-31 |
DE3320339A1 (de) | 1984-01-19 |
SE459816B (sv) | 1989-08-07 |
SE8303068L (sv) | 1983-12-05 |
US4703005A (en) | 1987-10-27 |
JPS592689A (ja) | 1984-01-09 |
GB2124232B (en) | 1985-12-18 |
GB8315278D0 (en) | 1983-07-06 |
NL189208B (nl) | 1992-09-01 |
FR2528070A1 (fr) | 1983-12-09 |
CA1202258A (en) | 1986-03-25 |
NL189208C (nl) | 1993-02-01 |
GB2124232A (en) | 1984-02-15 |
NL8301986A (nl) | 1984-01-02 |
SE8303068D0 (sv) | 1983-05-31 |
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