EP0848754A1 - Procedes de clonage servant a obtenir des proteines de soie d'araignee extremement resistantes - Google Patents

Procedes de clonage servant a obtenir des proteines de soie d'araignee extremement resistantes

Info

Publication number
EP0848754A1
EP0848754A1 EP96932937A EP96932937A EP0848754A1 EP 0848754 A1 EP0848754 A1 EP 0848754A1 EP 96932937 A EP96932937 A EP 96932937A EP 96932937 A EP96932937 A EP 96932937A EP 0848754 A1 EP0848754 A1 EP 0848754A1
Authority
EP
European Patent Office
Prior art keywords
dna
silk
protein
spider
primers
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.)
Withdrawn
Application number
EP96932937A
Other languages
German (de)
English (en)
Inventor
Richard M. Basel
Glenn R. Elion
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of EP0848754A1 publication Critical patent/EP0848754A1/fr
Withdrawn legal-status Critical Current

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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
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
    • C07K14/43518—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from spiders

Definitions

  • This invention relates to novel methods of producing DNA fragments encoding for spider silk proteins.
  • the present invention also relates to the DNA sequences encoding the spider silk proteins.
  • This invention still further relates to novel methods of producing spider silk proteins using the above-described DNA sequences.
  • the invention also relates to methods of purifying these spider silk proteins and manufacturing fibers and films from them.
  • Nephila clavipes dragline silk was taken by Xu et al. (Proc. Natl. Acad. Sci. 87:7120, 1990) .
  • Xu et al. ascertained a portion of the repetitive sequence of a spider dragline silk from a partial clone. Although this repeating unit encoded for up to 34 amino acids, it was not exactly conserved as the sequence had deletions and changes in some of the repeats. Nevertheless, Xu et al. discovered two important areas in the sequence -- repetitive regions which give spider silk some of their properties and a non-repetitive (carboxy) region. Hinman and Lewis (J: Biol. Chem.
  • a second primer site is created at the unknown end of the DNA using a ligation cassette.
  • a second primer site is created at the unknown end of the DNA using a terminal transferase to make a primer site selected from the group con ⁇ isting of poly dT, poly dA, poly dG and poly dC.
  • This multimerizat. on process further comprises the steps of (iv) selecting a second pair of different DNA primers, at least one of the second pair of DNA primers being different than both of the sequences of the first pair of DNA primers, and at least one of the second pair of DNA primers being represented by the sequences (i) - (xxvi) ; (v) producing a second DNA fragment by repetitively combining the second pair of DNA primers with melted target DNA and incubating the combined DNA primers and target DNA with nucleotides and a DNA polymerase having proofreading ability to produce the second DNA fragment, the second DNA fragment being different than the first DNA fragment and also being complementary to the target DNA, the second DNA fragment being at least 2 Kb; (vi) restricting the first and ⁇ econd DNA fragment ⁇ ; and (vii) recombining the restricted portions of the first and second DNA fragments into a multimerized DNA, the multimerized DNA encoding spider silk protein and being at lea ⁇ t 4 Kb in length.
  • primer sequences (i) - (xx) Some of the primers used are disclosed above as primer sequences (i) - (xx) . Although these primers were also tried by Beckwith & Arcidiacono, the present inventors are the first to produce spider silk protein up to 2 Kb in length using a two primer PCR cloning system. The present inventors were also able to produce spider silk proteins with higher Kbs by the claimed cDNA and ⁇ ingle site cloning methods de ⁇ cribed below.
  • primer (iii) GCATGCACGCATGGTGCATGGATGC
  • primer (ii) GGCGAATTCACCCTAGGGCTTGATAAACTGATTGAC primer (iii) was made from the peptide sequence 4 described by Mello et al. , Silk Polymers, ACS, Symposium, Ser 544 (1994) .
  • Primer (ii) was made as described in Example 1 above.
  • PCR mix 5 ⁇ l 10X Takara LA PCR buffer; 5 ⁇ l Takara dNTP mix; 1 ⁇ l primer (iii) (2 ⁇ M) ; 1 ⁇ l primer (ii) (2 ⁇ M) ; l ⁇ l Takara Ex Taq with proofreading activity; 1 ⁇ l spider genomic DNA; water to a total of 50 ⁇ l; and 50 ⁇ l mineral oil.
  • the Takara LA PCR buffer, dNTP mix, and Takara Ex Taq were supplied with a Takara Roll kit distributed by Panvera Corp., 565 Science Dr., Madison, WI 53711. PCR cycler conditions were as follows: initial dwell 94°C.
  • Positive transformant ⁇ were assayed for insertion by checking the size of insertion with a 1% agarose gel.
  • the positive inserts were then tested for the correct insert by using PCR and poly d(T) 20 primer.
  • the positives were also tested by the antibody methods discu ⁇ ed below.
  • the positives passing the antibody tests for large mRNA were tested using SDS electrophoresis gels and found to give three different proteins also proving multiple start sites.
  • One protein was slightly larger than the 2 Kb piece and the other two proteins were slightly shorter than native ⁇ pider silk dragline protein. It was difficult, however, to get these high molecular weight proteins to stain with a Western stain, but this was also true with the native proteins.
  • the 2 Kb inserts were the longest spider silk pieces cloned. Because of this, it was theorized that a different technique would be required to make larger fragments. It was considered necessary that the technique obtain additional sequence information from parts of the protein coding towards the amino end because, with the available information from the protein sequencing, larger fragments were not produced. Although the 2 Kb piece was over 40% of full length, multimerization was considered necessary to increase strength characteristics -- as strength generally varies with the size of the ⁇ ilk polymer. Therefore, the inventors wanted to multimerize the 2 Kb insert to make a larger protein than the natural gene.
  • Bacillu ⁇ expre ⁇ sion system ⁇ including Ex. subtilis sy ⁇ terns can also be used. These bacteria have the advantage of good secretion by the host, which results in less processing steps and processing costs. Although an expression cassette might be used, it has been found unneces ⁇ ary with the vector host system ⁇ studied thus far.
  • One phagemid that can act as an EX . coli and Bacillus shuttle vector is pTZ18R which can be obtained from Pharmacia (Piscataway, NJ) .
  • the purification of silk protein from the fermentation media can be accomplished by a two step process.
  • the bacterial cells and precipitated protein can be removed by continuous centrifugation.
  • the remaining material present in the fermentation broth can be separated by ultrafiltration since most of the protein above a molecular weight of 80,000 is silk.
  • the protein silk streams from the continuous centrifugation and ultrafiltration procedures can then be combined.
  • the bulk of the remaining proteins can be found in the bacterial membranes.
  • By rupturing the bacterial cells using ultrasound the cells are opened and the ⁇ ilk protein in them i ⁇ removed.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Insects & Arthropods (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Chemical Or Physical Treatment Of Fibers (AREA)
  • Artificial Filaments (AREA)
  • Woven Fabrics (AREA)

Abstract

L'invention concerne de procédés de production de fragments d'ADN codant des protéines de soie provenant d'araignées. Elle concerne également les séquences d'ADN codant lesdites protéines, ainsi que des procédés de production de protéines de soie d'araignée au moyen des séquences d'ADN mentionnées ci-dessus. Ces procédés de clonage et de production de protéines peuvent être appliqués à toutes les araignées productrices de soie. Les clones obtenus au moyen de ces procédés produisent des quantités exploitables commercialement de protéines de soie à poids moléculaire élevé. Etant donné que la soie produite à partir de ces protéines présente des propriétés supérieures de résistance, ces protéines de soie clonées sont d'une importance considérable sur le plan industriel.
EP96932937A 1995-08-22 1996-08-22 Procedes de clonage servant a obtenir des proteines de soie d'araignee extremement resistantes Withdrawn EP0848754A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US51769495A 1995-08-22 1995-08-22
US517694 1995-08-22
PCT/US1996/013767 WO1997008315A1 (fr) 1995-08-22 1996-08-22 Procedes de clonage servant a obtenir des proteines de soie d'araignee extremement resistantes

Publications (1)

Publication Number Publication Date
EP0848754A1 true EP0848754A1 (fr) 1998-06-24

Family

ID=24060849

Family Applications (1)

Application Number Title Priority Date Filing Date
EP96932937A Withdrawn EP0848754A1 (fr) 1995-08-22 1996-08-22 Procedes de clonage servant a obtenir des proteines de soie d'araignee extremement resistantes

Country Status (7)

Country Link
EP (1) EP0848754A1 (fr)
JP (1) JPH11511325A (fr)
CN (1) CN1200145A (fr)
AU (1) AU7152996A (fr)
BR (1) BR9612625A (fr)
IL (1) IL123398A0 (fr)
WO (1) WO1997008315A1 (fr)

Families Citing this family (84)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2774588B1 (fr) 1998-02-11 2000-05-05 Oreal Composition cosmetique ou dermatologique contenant au moins une proteine de soie d'arachnides naturelle, recombinante ou un analogue
US7157615B2 (en) * 1998-03-17 2007-01-02 Nexia Biotechnologies, Inc. Production of biofilaments in transgenic animals
AU2001231451A1 (en) * 2000-02-03 2001-08-14 Nexia Biotechnologies, Inc. Surgical sutures containing spider silk
US6608242B1 (en) 2000-05-25 2003-08-19 E. I. Du Pont De Nemours And Company Production of silk-like proteins in plants
EP1287139B1 (fr) * 2000-06-09 2010-08-25 Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK) Proteines de soie d'araignee synthetiques et leur expression dans des plantes transgeniques
CA2351274C (fr) * 2000-06-29 2011-04-26 Fiona F. Hunter Proteine isolee de soie de cocon de simulium vittatum et acides nucleiques encodant une telle proteine
US20110009960A1 (en) 2001-11-16 2011-01-13 Allergan, Inc. Prosthetic fabric structure
GB2399820A (en) * 2002-01-11 2004-09-29 Nexia Biotech Inc Recovery of biofilament proteins from biological fluids
CA2525994C (fr) 2002-06-24 2012-10-16 Tufts University Biomateriaux a base de soie et leurs methodes d'utilisation
DK2374919T3 (da) 2003-03-11 2013-08-05 Allergan Inc Biokompatibelt helingsunderstøttende silkestof
CA2562415C (fr) 2003-04-10 2015-10-27 Tufts University Solution aqueuse concentree de fibroine
NZ567202A (en) * 2005-10-05 2012-02-24 Commw Scient Ind Res Org Silk proteins from Hyhemonptera species containing heptad repeats that form coiled coil tertiary structures
AU2006333611B2 (en) 2005-12-30 2012-04-26 Spiber Technologies Ab Spider silk proteins and methods for producing spider silk proteins
WO2008118133A2 (fr) 2006-09-26 2008-10-02 Trustees Of Tufts College Microsphères de soie pour l'encapsulation et la libération contrôlée
WO2008106485A2 (fr) 2007-02-27 2008-09-04 Trustees Of Tufts College Organes de soie produits par génie tissulaire
WO2009023615A1 (fr) 2007-08-10 2009-02-19 Trustees Of Tufts College Compositions de soie tubulaires et procédés d'utilisation de celles-ci
CN101970023A (zh) * 2008-02-07 2011-02-09 塔夫茨大学信托人 三维的丝羟基磷灰石组合物
JP5896743B2 (ja) 2008-10-09 2016-03-30 タフツ ユニバーシティー／トラスティーズ オブ タフツ カレッジ グリセロールを含有する改変された絹フィルム
WO2010057142A2 (fr) 2008-11-17 2010-05-20 Trustees Of Tufts College Modification de surface de matrices de fibroïne de soie avec du poly(éthylène glycol) utilisable comme barrière anti-adhérence et matériau anti-thrombose
US9308070B2 (en) 2008-12-15 2016-04-12 Allergan, Inc. Pliable silk medical device
US20130245757A1 (en) 2011-11-04 2013-09-19 Allergan, Inc. Method and device for improved soft tissue surgery
US20120070427A1 (en) 2009-06-01 2012-03-22 Trustees Of Tufts College Vortex-induced silk fibroin gelation for encapsulation and delivery
AU2010307268B2 (en) 2009-07-20 2015-05-14 Tufts University/Trustees Of Tufts College All-protein implantable, resorbable reflectors
CA2773956A1 (fr) 2009-09-11 2011-03-17 Allergan, Inc. Dispositif prothetique et son procede de fabrication
WO2011109691A2 (fr) 2010-03-05 2011-09-09 Trustees Of Tufts College Compositions ionomères à base de soie
EP2559101B1 (fr) 2010-04-12 2016-02-24 Tufts University Composants électroniques de soie
KR20130138763A (ko) 2010-08-30 2013-12-19 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 고강도 키틴 복합 재료 및 그 제조 방법
JP2013542181A (ja) 2010-09-03 2013-11-21 タフツ ユニバーシティー／トラスティーズ オブ タフツ カレッジ プラズモンナノ粒子ドープ絹材料
JP5691052B2 (ja) * 2010-09-10 2015-04-01 岡本株式会社 組換え生物及び組換え生物により作られるタンパク質
EP2622663A4 (fr) 2010-09-27 2014-12-24 Tufts University Trustees Of Tufts College Matériaux piézoélectriques à base de soie
CA2815285C (fr) 2010-10-19 2019-12-31 Trustees Of Tufts College Micro-aiguilles a base de fibroine de soie et procedes pour les fabriquer
WO2012145652A1 (fr) 2011-04-20 2012-10-26 Trustees Of Tufts College Revêtements de soie dynamiques pour dispositifs implantables
BR112013027057A2 (pt) 2011-04-21 2020-08-11 Trustees Of Tufts College composições e métodos para a estabilização de agentes ativos
US20140370094A1 (en) 2011-11-08 2014-12-18 Tufts University Silk-based scaffold platform for engineering tissue constructs
BR112014011188B1 (pt) 2011-11-09 2020-05-19 Trustees Of Tufts College composição injetável para utilização na reparação ou aumento de tecido em indivíduo e dispositivo de entrega compreendendo a referida composição
AU2012335122B2 (en) 2011-11-09 2016-09-22 Trustees Of Tufts College Injectable silk fibroin foams and uses thereof
US10126467B2 (en) 2011-12-05 2018-11-13 Tufts University Signal enhancement by silk photonic crystals
WO2013102193A1 (fr) 2011-12-29 2013-07-04 Trustees Of Tufts College Fonctionnalisation de biomatériaux pour commander la régénération et des réponses à une inflammation
ES2856873T3 (es) 2012-02-06 2021-09-28 Childrens Medical Center Biomaterial multicapa para la regeneración de tejidos y la cicatrización de las heridas
NL1039495C2 (nl) * 2012-03-27 2013-09-30 Jalila Essaidi Werkwijze ter vervaardiging van een huidsubstituut op basis van spinnenzijde alsmede draagstructuur vervaardigd onder toepassing van deze werkwijze en draagstructuur met doorgroeiende levende huid.
CN104602713A (zh) 2012-04-13 2015-05-06 塔夫茨大学信托人 用于制备丝微球的方法和组合物
EP2841108A4 (fr) 2012-04-25 2015-11-25 Tufts College Microsphères de soie et procédés de lubrification de surface
WO2014012101A1 (fr) 2012-07-13 2014-01-16 Trustees Of Tufts College Compactage de poudre de soie pour fabrication de constructions à résistance mécanique et rigidité élevées
WO2014035798A1 (fr) 2012-08-30 2014-03-06 Sanofi Lyogels de soie pour la libération prolongée de produits thérapeutiques protéiques et procédés de fabrication et d'utilisation associés
EP2925822A4 (fr) 2012-11-27 2016-10-12 Univ Tufts Encres à base de biopolymères et leur utilisation
JP6317425B2 (ja) 2013-03-15 2018-04-25 パセオン ソフトジェルズ インコーポレイティド シルク系カプセル剤
US10464361B2 (en) 2013-03-15 2019-11-05 Tufts University Silk water lithography
US11376329B2 (en) 2013-03-15 2022-07-05 Trustees Of Tufts College Low molecular weight silk compositions and stabilizing silk compositions
KR102457668B1 (ko) 2013-03-15 2022-10-21 트러스티즈 오브 터프츠 칼리지 저분자량 실크 조성물 및 안정화 실크 조성물
WO2014197093A2 (fr) 2013-03-15 2014-12-11 Tufts University Formation de nanomotif à base d'eau
CN103194068B (zh) * 2013-04-08 2015-12-23 浙江大学 一种稳定丝素蛋白溶液的方法
WO2014176458A2 (fr) 2013-04-24 2014-10-30 Trustees Of Tufts College Dispositifs pour anastomose en biopolymère biorésorbable
WO2015048527A1 (fr) 2013-09-27 2015-04-02 Tufts University Hydrogels de soie optiquement transparents
US10925999B2 (en) 2013-10-08 2021-02-23 Trustees Of Tufts College Tunable covalently crosslinked hydrogels and methods of making the same
US10513802B2 (en) 2013-11-08 2019-12-24 Tufts University Peptide-based nanofibrillar materials
US10271561B2 (en) 2014-03-07 2019-04-30 Tufts University Biopolymer-based preservation of perishable products
WO2015178466A1 (fr) * 2014-05-21 2015-11-26 味の素株式会社 Procédé de production de protéines de type fibroïne
WO2016028797A1 (fr) 2014-08-18 2016-02-25 Allergan, Inc. Dispositif médical en soie pliable
PE20171791A1 (es) 2014-12-02 2017-12-28 Silk Therapeutics Inc Prendas de vestir de seda y productos de alto rendimiento y metodos para elaborarla
WO2016100721A1 (fr) 2014-12-17 2016-06-23 Tufts University Mousses souple et injectable à base d'hydroxyapatite et de soie pour réparation dentaire et ostéochondrale
US10874742B2 (en) 2015-03-12 2020-12-29 Tufts University Shape memory silk materials
CA2995233C (fr) 2015-07-20 2024-01-02 Tufts University Tubes d'oreille de soie biodegradables
JP2018527978A (ja) 2015-07-28 2018-09-27 トラスティーズ オブ タフツ カレッジ ポリペプチドモノリス
CN105231565B (zh) * 2015-10-20 2018-01-16 杭州华水布艺有限公司 一种含冰凉材料的防刺防护面料
US11833272B2 (en) 2015-12-17 2023-12-05 Trustees Of Tufts College Silk-fibroin hydrogels, methods of forming, and uses thereof
WO2017106631A1 (fr) 2015-12-18 2017-06-22 Tufts University Système de purification de solutions de soie, système de concentration et procédés associés
US11643444B2 (en) 2016-05-04 2023-05-09 Trustees Of Tufts College Silk nanofibrils and uses thereof
WO2018006037A1 (fr) 2016-07-01 2018-01-04 Trustees Of Tufts College Peau artificielle innervée
WO2018026853A1 (fr) 2016-08-01 2018-02-08 Trustees Of Tufts College Fabrication assistée par tension mécanique biomimétique d'une architecture nanofibrillaire
EP3532093A1 (fr) 2016-09-19 2019-09-04 Vaxess Technologies, Inc. Formulations de vaccin présentant une stabilité accrue
WO2018081159A1 (fr) 2016-10-24 2018-05-03 Trustees Of Tufts College Compositions multicouches biomimétiques
US10857262B2 (en) 2016-10-31 2020-12-08 Sofregen Medical, Inc. Compositions comprising low molecular weight silk fibroin fragments and plasticizers
WO2018136754A1 (fr) 2017-01-20 2018-07-26 Massachusetts Institute Of Technology Micro-dépôts polymères injectables pour administration locale contrôlée de médicament
US20200306163A1 (en) * 2017-09-27 2020-10-01 Evolved By Nature, Inc. Materials comprising recombinant silk and methods of preparing the same
US11419947B2 (en) 2017-10-30 2022-08-23 Massachusetts Institute Of Technology Layer-by-layer nanoparticles for cytokine therapy in cancer treatment
AU2019247655A1 (en) 2018-04-03 2020-10-01 Vaxess Technologies, Inc. Microneedle comprising silk fibroin applied to a dissolvable base
WO2020023906A2 (fr) 2018-07-27 2020-01-30 Vaxess Technologies, Inc. Dispositifs de prélèvement d'échantillons biologiques à base de polymère et leurs utilisations
JP7737142B2 (ja) * 2018-11-28 2025-09-10 ボルト スレッズ インコーポレイテッド クモ糸タンパク質のアルカリ精製法
CN110106571A (zh) * 2019-04-09 2019-08-09 商文辉 一种蛛网纺织面料及其制备方法
EP3975841A1 (fr) 2019-05-30 2022-04-06 Massachusetts Institute of Technology Micro-aiguilles d'hydrogel fonctionnalisées par de l'acide nucléique peptidique pour l'échantillonnage et la détection d'acides nucléiques fluides interstitiels
EP4218843A3 (fr) * 2019-09-16 2023-08-23 Bolt Threads, Inc. Procédés pour isoler des protéines de soie d'araignée par l'intermédiaire d'une solubilisation à cisaillement élevé
EP4045124A4 (fr) 2019-10-15 2024-01-24 Sofregen Medical, Inc. Dispositifs de distribution pour l'administration et procédés de distribution de compositions
JP7670355B2 (ja) 2019-12-06 2025-04-30 トラスティーズ オブ タフツ カレッジ 組織再生多剤カクテル及びその送達のための装置
US20240104583A1 (en) 2022-09-26 2024-03-28 Massachusetts Institute Of Technology Integrating Biopolymer Design with Physical Unclonable Functions for Anticounterfeiting and Product Traceability

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU7691191A (en) * 1990-04-19 1991-11-11 United States Of America, As Represented By The Secretary Of The Army, The Recombinant spider silk proteins through genetic engineering
DE69131969T2 (de) * 1990-04-20 2000-06-15 The University Of Wyoming, Laramie Für Spinnenseideprotein kodierende DNS, DNS enthaltender Vektor, transformierte Zelle und Produkte davon

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9708315A1 *

Also Published As

Publication number Publication date
BR9612625A (pt) 1999-06-01
CN1200145A (zh) 1998-11-25
WO1997008315A1 (fr) 1997-03-06
AU7152996A (en) 1997-03-19
IL123398A0 (en) 1998-09-24
JPH11511325A (ja) 1999-10-05

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