LT4000B - Expression module, transformed host cell, process for preparing serine protease, method of transformation, dna sequence, plasmid, oligonucleotide - Google Patents
Expression module, transformed host cell, process for preparing serine protease, method of transformation, dna sequence, plasmid, oligonucleotide Download PDFInfo
- Publication number
- LT4000B LT4000B LTIP1820A LTIP1820A LT4000B LT 4000 B LT4000 B LT 4000B LT IP1820 A LTIP1820 A LT IP1820A LT IP1820 A LTIP1820 A LT IP1820A LT 4000 B LT4000 B LT 4000B
- Authority
- LT
- Lithuania
- Prior art keywords
- bacillus
- host cell
- serine protease
- dna
- host
- Prior art date
Links
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
-
- 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
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
-
- 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
- C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N15/75—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Bacillus
-
- 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
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
-
- 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)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
- C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP87200358 | 1987-02-27 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| LTIP1820A LTIP1820A (en) | 1995-08-25 |
| LT4000B true LT4000B (en) | 1996-06-25 |
Family
ID=8197584
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| LTIP1820A LT4000B (en) | 1987-02-27 | 1994-01-28 | Expression module, transformed host cell, process for preparing serine protease, method of transformation, dna sequence, plasmid, oligonucleotide |
Country Status (19)
| Country | Link |
|---|---|
| US (1) | US5217878A (da) |
| JP (1) | JP3026567B2 (da) |
| KR (1) | KR0127560B1 (da) |
| CN (1) | CN88101681A (da) |
| AT (2) | ATE181110T1 (da) |
| AU (1) | AU620026B2 (da) |
| BG (1) | BG49049A3 (da) |
| BR (1) | BR8805647A (da) |
| CA (1) | CA1339100C (da) |
| DE (2) | DE3856331T2 (da) |
| DK (2) | DK175738B1 (da) |
| FI (3) | FI105483B (da) |
| HU (1) | HU213220B (da) |
| LT (1) | LT4000B (da) |
| LV (1) | LV10307B (da) |
| NO (1) | NO884423D0 (da) |
| PT (1) | PT86864B (da) |
| RU (1) | RU2023723C1 (da) |
| WO (1) | WO1988006624A2 (da) |
Families Citing this family (52)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1988006624A2 (en) * | 1987-02-27 | 1988-09-07 | Gist-Brocades N.V. | Molecular cloning and expression of genes encoding proteolytic enzymes |
| DK6488D0 (da) * | 1988-01-07 | 1988-01-07 | Novo Industri As | Enzymer |
| GB2214508A (en) * | 1988-01-27 | 1989-09-06 | Bayer Ag | Plasmid vector for the efficient expression of foreign genes fused with newly conceived transcriptional and translational signal sequences. |
| US6287841B1 (en) * | 1988-02-11 | 2001-09-11 | Genencor International, Inc. | High alkaline serine protease |
| WO1989010976A1 (en) * | 1988-05-05 | 1989-11-16 | The Public Health Research Institute Of The City O | Protease-deficient gram-positive bacteria and their use as host organisms for the production of recombinant products |
| US5665587A (en) * | 1989-06-26 | 1997-09-09 | Novo Nordisk A/S | Modified subtilisins and detergent compositions containing same |
| JP3046344B2 (ja) * | 1990-10-24 | 2000-05-29 | 塩野義製薬株式会社 | 新規プロテアーゼ |
| US5482849A (en) * | 1990-12-21 | 1996-01-09 | Novo Nordisk A/S | Subtilisin mutants |
| DK58391D0 (da) * | 1991-04-03 | 1991-04-03 | Novo Nordisk As | Hidtil ukendte proteaser |
| EP0571014B1 (en) * | 1992-05-18 | 2004-03-31 | Genencor International, Inc. | Bacteria producing alkaline proteases, and production of these alkaline proteases |
| US6436690B1 (en) * | 1993-09-15 | 2002-08-20 | The Procter & Gamble Company | BPN′ variants having decreased adsorption and increased hydrolysis wherein one or more loop regions are substituted |
| US6440717B1 (en) * | 1993-09-15 | 2002-08-27 | The Procter & Gamble Company | BPN′ variants having decreased adsorption and increased hydrolysis |
| US6599730B1 (en) * | 1994-05-02 | 2003-07-29 | Procter & Gamble Company | Subtilisin 309 variants having decreased adsorption and increased hydrolysis |
| US5847135A (en) * | 1994-06-17 | 1998-12-08 | Vertex Pharmaceuticals, Incorporated | Inhibitors of interleukin-1β converting enzyme |
| US5831004A (en) * | 1994-10-27 | 1998-11-03 | Affymax Technologies N.V. | Inhibitors of metalloproteases, pharmaceutical compositions comprising same and methods of their use |
| US5646044A (en) * | 1995-03-02 | 1997-07-08 | Henkel Kommanditgesellschaft Auf Aktien | Expression systems for the production of target proteins in bacillus |
| US6455295B1 (en) * | 1995-03-08 | 2002-09-24 | The Procter & Gamble Company | Subtilisin Carlsberg variants having decreased adsorption and increased hydrolysis |
| IL117350A0 (en) * | 1995-03-09 | 1996-07-23 | Procter & Gamble | Proteinase k variants having decreased adsorption and increased hydrolysis |
| US6475765B1 (en) * | 1995-03-09 | 2002-11-05 | Procter & Gamble Company | Subtilisin DY variants having decreased adsorption and increased hydrolysis |
| EP0932667B1 (en) | 1996-11-04 | 2008-10-01 | Novozymes A/S | Subtilase variants and compositions |
| US5766871A (en) * | 1996-11-27 | 1998-06-16 | Food Industry Research And Development Institute | Screening and characterization of glutaryl-7-aminocephalosporanic acid acylase |
| US6485957B1 (en) * | 1999-04-30 | 2002-11-26 | Ortho-Mcneil Pharmaceutical, Inc. | DNA encoding the human serine protease EOS |
| US6420157B1 (en) * | 1999-04-30 | 2002-07-16 | Ortho-Mcneil Pharmaceutical, Inc. | Zymogen activation system |
| US6458564B1 (en) | 1999-08-31 | 2002-10-01 | Ortho-Mcneil Pharmaceutical, Inc. | DNA encoding the human serine protease T |
| US6426199B1 (en) | 1999-08-31 | 2002-07-30 | Ortho-Mcneil Pharmaceutical, Inc. | Dna |
| CN1312370A (zh) * | 2000-03-07 | 2001-09-12 | 上海博德基因开发有限公司 | 一种新的多肽——人atp依赖的丝氨酸蛋白水解酶11和编码这种多肽的多核苷酸 |
| US6777218B1 (en) * | 2000-03-14 | 2004-08-17 | Novozymes A/S | Subtilase enzymes having an improved wash performance on egg stains |
| JP2004508011A (ja) * | 2000-04-03 | 2004-03-18 | マキシジェン, インコーポレイテッド | スブチリシン変異体 |
| DK1495128T3 (da) | 2002-03-29 | 2014-08-11 | Genencor Int | Forstærket proteinekspression i Bacillus |
| US20100129862A1 (en) * | 2003-05-12 | 2010-05-27 | Jones Brian E | Novel lipolytic Enzyme lip1 |
| EP1625208A4 (en) * | 2003-05-12 | 2006-10-18 | Genencor Int | NEW LIPOLYTIC ENZYME LIP2 |
| WO2004101760A2 (en) * | 2003-05-12 | 2004-11-25 | Genencor International, Inc. | Novel lipolytic enzyme elip |
| CA2539423C (en) | 2003-09-19 | 2011-09-06 | Ics Holdings, Inc | Preparation of an edible product from dough |
| US8535927B1 (en) * | 2003-11-19 | 2013-09-17 | Danisco Us Inc. | Micrococcineae serine protease polypeptides and compositions thereof |
| US7985569B2 (en) | 2003-11-19 | 2011-07-26 | Danisco Us Inc. | Cellulomonas 69B4 serine protease variants |
| WO2006033668A2 (en) * | 2004-04-09 | 2006-03-30 | Genencor International, Inc. | Pcka modifications and enhanced protein expression in bacillus |
| WO2005123915A1 (en) * | 2004-06-21 | 2005-12-29 | Novozymes A/S | Stably maintained multiple copies of at least two orf in the same orientation |
| US7202074B2 (en) * | 2004-07-22 | 2007-04-10 | University Of Chile | Protein and nucleic acid sequence encoding a KRILL-derived cold adapted trypsin-like activity enzyme |
| EP2129779B2 (en) | 2007-03-12 | 2018-12-26 | Danisco US Inc. | Modified proteases |
| DK2152732T3 (da) | 2007-05-10 | 2012-06-04 | Danisco Us Inc | Modificeret sekretionssystem til at øge ekspression af polypeptider i bakterier |
| US8759065B2 (en) | 2007-08-06 | 2014-06-24 | University Of Chile | Protein and DNA sequence encoding a cold adapted subtilisin-like activity |
| AU2008348270A1 (en) | 2007-12-21 | 2009-07-30 | Danisco Us Inc. | Enhanced protein production in bacillus |
| FI123122B (fi) | 2009-02-16 | 2012-11-15 | Optogear Oy | Laitteisto lasimateriaalin valmistamiseksi |
| AR076312A1 (es) | 2009-04-24 | 2011-06-01 | Danisco Us Inc | Proteasas con regiones pro modificadas |
| AR076941A1 (es) | 2009-06-11 | 2011-07-20 | Danisco Us Inc | Cepa de bacillus para una mayor produccion de proteina |
| US20140227738A1 (en) * | 2011-09-22 | 2014-08-14 | Novozymes A/S | Polypeptides Having Protease Activity and Polynucleotides Encoding Same |
| CN103013960A (zh) * | 2012-12-21 | 2013-04-03 | 青岛蔚蓝生物集团有限公司 | 一种碱性蛋白酶及其重组表达工程菌 |
| EP3089991B1 (en) | 2013-12-31 | 2019-08-28 | Danisco US Inc. | Enhanced protein expression |
| KR102588719B1 (ko) | 2014-12-16 | 2023-10-12 | 다니스코 유에스 인크. | 향상된 단백질 발현 |
| CN107278230B (zh) | 2014-12-19 | 2021-10-29 | 丹尼斯科美国公司 | 增强的蛋白质表达 |
| AU2018262177B2 (en) * | 2017-05-05 | 2023-05-18 | Bioecho Life Sciences Gmbh | Rapid purification of high quality nucleic acids from biological samples |
| US12139715B2 (en) | 2018-06-19 | 2024-11-12 | Seoul National University R&Db Foundation | Gene expression cassette capable of initiating translation after completion of transcription for producing high-quality recombinant protein in bacteria |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3674643A (en) | 1967-11-10 | 1972-07-04 | Novo Terapeutisk Labor As | Preparation of proteolytic enzymes having maximum activity at high alkalinity |
| US3723250A (en) | 1967-10-03 | 1973-03-27 | Novo Terapeutisk Labor As | Proteolytic enzymes, their production and use |
| US4480037A (en) | 1982-02-08 | 1984-10-30 | Showa Denko Kabushiki Kaisha | Alkaline protease and preparation method thereof |
| US4480043A (en) | 1980-05-20 | 1984-10-30 | Biomerieux | Process for the preparation of toxoplasmas for the diagnosis of toxoplasmosis, and preparations thus obtained |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1519148A (en) * | 1974-11-19 | 1978-07-26 | Gist Brocades Nv | Compositions of matter |
| ATE48156T1 (de) * | 1982-11-01 | 1989-12-15 | Miles Inc | Verfahren zur heterologen klonierung eines gens in einem bacillus-mikroorganismus. |
| NZ208612A (en) * | 1983-06-24 | 1991-09-25 | Genentech Inc | Method of producing "procaryotic carbonyl hydrolases" containing predetermined, site specific mutations |
| DE3480411D1 (en) * | 1983-07-06 | 1989-12-14 | Gist Brocades Nv | Molecular cloning and expression in industrial microorganism species |
| EP0138075A1 (en) * | 1983-09-16 | 1985-04-24 | Synergen Associates, Inc. | A method for the generation and detection of enzyme variants with enhanced themostability and activity |
| US4828994A (en) * | 1984-09-21 | 1989-05-09 | Genex Corporation | Bacillus strains with reduced extracellular protease levels |
| FR2582316B1 (fr) * | 1985-05-22 | 1988-12-02 | Centre Nat Rech Scient | Vecteurs d'expression de l'a-amylase dans bacillus subtilis, souches obtenues et procede de preparation d'a-amylase |
| DE3527913A1 (de) * | 1985-08-03 | 1987-02-12 | Henkel Kgaa | Alkalische protease, verfahren zur herstellung von hybridvektoren und genetisch transformierte mikroorganismen |
| EP0254735B2 (en) * | 1986-01-15 | 1998-06-17 | Amgen Inc. | METHOD FOR PRODUCTION OF THERMALLY STABLE AND pH STABLE SUBTILISIN ANALOGS |
| IE65767B1 (en) * | 1986-04-30 | 1995-11-15 | Genencor Int | Non-human carbonyl hydrolase mutants DNA sequences and vectors encoding same and hosts transformed with said vectors |
| WO1988006624A2 (en) * | 1987-02-27 | 1988-09-07 | Gist-Brocades N.V. | Molecular cloning and expression of genes encoding proteolytic enzymes |
| ES2135386T3 (es) * | 1987-02-27 | 1999-11-01 | Genencor Int | Transformacion de cepas de bacillus alcalofilas. |
-
1988
- 1988-02-26 WO PCT/NL1988/000007 patent/WO1988006624A2/en not_active Ceased
- 1988-02-26 AU AU13980/88A patent/AU620026B2/en not_active Expired
- 1988-02-26 BR BR888805647A patent/BR8805647A/pt not_active IP Right Cessation
- 1988-02-26 PT PT86864A patent/PT86864B/pt not_active IP Right Cessation
- 1988-02-26 KR KR1019880701347A patent/KR0127560B1/ko not_active Expired - Fee Related
- 1988-02-26 JP JP63502472A patent/JP3026567B2/ja not_active Expired - Lifetime
- 1988-02-26 HU HU881832A patent/HU213220B/hu not_active IP Right Cessation
- 1988-02-27 CN CN198888101681A patent/CN88101681A/zh active Pending
- 1988-02-29 US US07/162,184 patent/US5217878A/en not_active Expired - Lifetime
- 1988-02-29 AT AT91203297T patent/ATE181110T1/de not_active IP Right Cessation
- 1988-02-29 DE DE3856331T patent/DE3856331T2/de not_active Expired - Lifetime
- 1988-02-29 CA CA000560126A patent/CA1339100C/en not_active Expired - Lifetime
- 1988-02-29 DE DE3856339T patent/DE3856339T2/de not_active Expired - Lifetime
- 1988-02-29 AT AT88200377T patent/ATE180016T1/de not_active IP Right Cessation
- 1988-10-05 NO NO884423A patent/NO884423D0/no unknown
- 1988-10-24 FI FI884904A patent/FI105483B/fi active IP Right Grant
- 1988-10-25 BG BG085831A patent/BG49049A3/xx unknown
- 1988-10-26 DK DK198805939A patent/DK175738B1/da not_active IP Right Cessation
- 1988-10-26 RU SU884356848A patent/RU2023723C1/ru active
-
1992
- 1992-11-27 FI FI925413A patent/FI106726B/fi active
-
1993
- 1993-12-29 LV LVP-93-1390A patent/LV10307B/en unknown
-
1994
- 1994-01-28 LT LTIP1820A patent/LT4000B/lt not_active IP Right Cessation
-
2000
- 2000-05-15 FI FI20001152A patent/FI119028B/fi not_active IP Right Cessation
-
2004
- 2004-08-20 DK DK200401262A patent/DK175852B1/da not_active IP Right Cessation
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3723250A (en) | 1967-10-03 | 1973-03-27 | Novo Terapeutisk Labor As | Proteolytic enzymes, their production and use |
| US3674643A (en) | 1967-11-10 | 1972-07-04 | Novo Terapeutisk Labor As | Preparation of proteolytic enzymes having maximum activity at high alkalinity |
| US4480043A (en) | 1980-05-20 | 1984-10-30 | Biomerieux | Process for the preparation of toxoplasmas for the diagnosis of toxoplasmosis, and preparations thus obtained |
| US4480037A (en) | 1982-02-08 | 1984-10-30 | Showa Denko Kabushiki Kaisha | Alkaline protease and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| CHANG S, COHEN SN.: "High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA.", MOL GEN GENET., 1979, pages 111 - 115, XP002092527, DOI: doi:10.1007/BF00267940 |
| MANIATIS T, ET AL.: "Molecular cloning" |
| P. VOROBJEVA ET AL.: "TRANSFORMATION OF BACILLUS MEGATERIUM PROTOPLASTS BY PLASMID DNA", FEMS MICROBIOLOGY LETTERS, 1980, pages 261 - 263, XP001313771 |
| PALVA I. ET AL.: "Nucleotide sequence of the promoter and NH2-terminal signal peptide region of the alpha-amylase gene from Bacillus amyloliquefaciens", GENE, 1981, pages 43 - 51, XP023542084, DOI: doi:10.1016/0378-1119(81)90103-7 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PC9A | Transfer of patents |
Free format text: GENENCOR INTERNATIONAL, INC.,4 CAMBRIDGE PLACE, 870 SOUTH WINTON ROAD, ROCHESTER NY 14618,US,970729 |
|
| MM9A | Lapsed patents |
Effective date: 19980128 |