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51.发明公开Method for producing human growth hormone 失效Methode zur Herstellung von menschlichen Wachstumshormonen。
公开(公告)号：EP0489711A2
公开(公告)日：1992-06-10
申请号：EP92101249.8
申请日：1985-08-09
CPC分类号： C12P21/06 , C07K14/555 , C07K14/61 , C12N9/48
摘要： The invention relates to a method of producing a human growth hormone having the amino acid sequence of naturally occurring human growth hormone which comprises
a) producing in a microbial host a first polypeptide which is characterized by the presence of one or more additional amino acids at the N-terminus of the amino acid sequence of naturally occurring human growth hormone;
b) contacting the first polypeptide so produced with an enzyme, preferably an aminopeptidase, so as to produce a second polypeptide having the amino acid sequence of naturally occurring human growth hormone; and
c) recovering the second polypeptide so produced.
A preferred microbial host is a bacterium, especially E. coli .摘要翻译： 本发明涉及一种生产具有天然存在的人生长激素的氨基酸序列的人生长激素的方法，其包括a）在微生物宿主中产生第一种多肽，其特征在于存在一种或多种另外的氨基酸 天然存在的人生长激素氨基酸序列的N末端; b）使如此产生的第一多肽与酶，优选氨基肽酶接触，以产生具有天然存在的人生长激素的氨基酸序列的第二多肽; 和c）回收如此制备的第二种多肽。 优选的微生物宿主是细菌，特别是大肠杆菌。
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52.发明公开A method of producing an enzymatically active polypeptide analog of human Cu/Zn SOD 失效生产人类CU / ZN SOD的酶促活性多肽模拟物的方法
公开(公告)号：EP0483113A3
公开(公告)日：1992-05-13
申请号：EP92101250.6
申请日：1985-08-26
发明人： Aviv, Haim , Bartfeld, Daniel , Gorecki, Marian , Hartman, Jacob R. , Kanner, Dov , Levanon, Avigdor , Locker-Giladi, Hilla , Oppenheim, Amos B.
CPC分类号： C12N9/0089 , A23K20/168 , A61K38/446 , C07K14/61 , C07K14/775 , C12N15/69 , C12N15/73
摘要： An improved vector which upon introduction into a suitable host containing the thermolabile repressor C I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P L O L from lambda bacteriophage; the N utilization site; a first restriction enzyme site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzme site for inserting the gene in phase with the ATG initiation codon; an origin of replication and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host or a fragment designated cI⁴³⁴ which includes the gene for the cI⁴³⁴ repressor protein and its associated promoter and operator. The distance between the 3' end of P L O L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Vectors of the invention may also include a T₁T₂ rRNA transcription termination sequence located 3' of the second restriction enzyme site. The inventive vectors may have origin of replications capable of autonomous production in the host of at least 400 constitutive copies. Plasmids have been constructed from the vectors and used to produce bovine, chicken, porcine and human growth hormones, human apolipoprotein E and human superoxide dismutase and analogs thereof in host cells. Such superoxide dismutase or analogs thereof may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and prevent neurological injury. Enzymatically active eucaryotic superoxide dismutase or an analog thereof may also be produced by an improved method of this invention which employs a production medium having a concentration of Cu⁺⁺ greater than about about 2ppm. The invention also concerns improved methods of recovering purified enzymatically active eucaryotic superoxide dismutase or analogs thereof.
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53.发明公开A method of producing an enzymatically active polypeptide analog of human Cu/Zn SOD 失效方法酶催化生物多磷酸盐分解酶Cu / Zn SOD herzustellen。
公开(公告)号：EP0483113A2
公开(公告)日：1992-04-29
申请号：EP92101250.6
申请日：1985-08-26
发明人： Aviv, Haim , Bartfeld, Daniel , Gorecki, Marian , Hartman, Jacob R. , Kanner, Dov , Levanon, Avigdor , Locker-Giladi, Hilla , Oppenheim, Amos B.
CPC分类号： C12N9/0089 , A23K20/168 , A61K38/446 , C07K14/61 , C07K14/775 , C12N15/69 , C12N15/73
摘要： An improved vector which upon introduction into a suitable host containing the thermolabile repressor C I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P L O L from lambda bacteriophage; the N utilization site; a first restriction enzyme site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzme site for inserting the gene in phase with the ATG initiation codon; an origin of replication and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host or a fragment designated cI⁴³⁴ which includes the gene for the cI⁴³⁴ repressor protein and its associated promoter and operator. The distance between the 3' end of P L O L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs.
Vectors of the invention may also include a T₁T₂ rRNA transcription termination sequence located 3' of the second restriction enzyme site. The inventive vectors may have origin of replications capable of autonomous production in the host of at least 400 constitutive copies.
Plasmids have been constructed from the vectors and used to produce bovine, chicken, porcine and human growth hormones, human apolipoprotein E and human superoxide dismutase and analogs thereof in host cells.
Such superoxide dismutase or analogs thereof may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and prevent neurological injury.
Enzymatically active eucaryotic superoxide dismutase or an analog thereof may also be produced by an improved method of this invention which employs a production medium having a concentration of Cu⁺⁺ greater than about about 2ppm.
The invention also concerns improved methods of recovering purified enzymatically active eucaryotic superoxide dismutase or analogs thereof.摘要翻译： 一种改进的载体，其在引入含有不耐热阻遏物CI的合适的宿主中时，使宿主能够实现所需基因的表达。 载体是双链DNA分子，其以5'至3'的顺序包括以下：来自λ噬菌体的启动子和操纵子PLOL; N利用场所; 此后第一个限制酶位点; 核糖体结合位点; ATG起始密码子或DNA，将所需基因插入载体后转化为ATG起始密码子; 用于将基因与ATG起始密码子相插入的第二限制性酶切位点; 复制起点和当载体存在于宿主中时表现出的可选择或可识别的表型性状相关的基因或称为cI4 <3>的片段，其包含cI4基因的基因， 3> 4阻抑蛋白及其相关启动子和操纵子。 PLOL的3'端与N使用位点的5'端之间的距离小于约80个碱基对。 N利用位点的3'末端与核糖体结合位点的5'末端之间的距离小于约300个碱基对。 本发明的载体还可以包含位于第二限制酶位点3'的T1T2 rRNA转录终止序列。 本发明的载体可以具有能够在宿主中自主生产至少400种组成型拷贝的重复起源。 已经从载体构建质粒，并用于在宿主细胞中产生牛，鸡，猪和人生长激素，人载脂蛋白E和人超氧化物歧化酶及其类似物。 这种超氧化物歧化酶或其类似物可用于催化超氧自由基的还原，减少再灌注损伤，延长分离的器官的存活时间并预防神经损伤。 酶促活性的真核超氧化物歧化酶或其类似物也可以通过本发明的改进方法生产，其使用浓度大于约2ppm的Cu 2+的生产培养基。 本发明还涉及回收纯化的酶促活性的真核超氧化物歧化酶或其类似物的改进方法。
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54.发明公开Expression plasmids containing the deo promoter, and bacterial hosts containing the plasmids 失效Den DEO-Promotor enthaltende Expressionsplasmide und dies Plasmide enthaltende bakterielle Wirte。
公开(公告)号：EP0303972A2
公开(公告)日：1989-02-22
申请号：EP88113071.0
申请日：1988-08-11
发明人： Fischer, Meir
CPC分类号： C12N9/0089 , C07K14/61 , C07K14/775 , C07K2319/00 , C07K2319/75 , C12N9/0065 , C12N15/70
摘要： Plasmids are provided which upon introduction into a suitable Escherichia coli host render the host capable of effecting expression of DNA encoding a desired naturally-occurring polypeptide or polypeptide analog thereof under the control of the deo P1-P2 promoter. Further plasmids are also provided which thermoinducibly direct expression of eucaryotic genes under the control of a λ P L promoter and a thermolabile repressor protein present on the same plasmid under the control of a deo P1 promoter. These plasmids have been inserted into various hosts, some of which contain DNA encoding the repressor protein.
Such plasmids may be used to produce polypeptides such as human copper/zinc superoxide dismutase, hMnSOD-hGPX fusion polypeptides, human manganese superoxide dismutase, human growth hormone, bovine growth, porcine growth, human growth hormone-apolipoprotein E fused polypeptides, apolipoprotein E and various fibronectin domains.摘要翻译： 提供质粒，其在引入合适的大肠杆菌宿主中时，使宿主能够在编码P1-P2启动子的控制下实现编码期望的天然存在的多肽或其多肽类似物的DNA的表达。 还提供了进一步的质粒，其在由λPL启动子和存在于相同质粒上的deo P1启动子控制下的热不稳定阻遏物蛋白质的温度诱导下直接表达真核基因。 这些质粒已被插入到各种宿主中，其中一些含有编码阻抑蛋白的DNA。 这样的质粒可用于产生多肽，例如人铜/锌超氧化物歧化酶，hMnSOD-hGPX融合多肽，人锰超氧化物歧化酶，人生长激素，牛生长，猪生长，人生长激素 - 载脂蛋白E融合多肽，载脂蛋白E和 各种纤连蛋白结构域。
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55.发明公开Human manganese superoxide dismutase and methods of treatment 失效人类锰过氧化物脱氧核糖核酸酶，其表达，回收人锰锰超氧化物歧化酶，人类锰超氧化物歧化酶模拟物或人类锰超氧化物歧化酶，用途，组合物和治疗方法
公开(公告)号：EP0284105A3
公开(公告)日：1989-01-25
申请号：EP88104880.5
申请日：1988-03-25
发明人： Hartman, Jacob R. , Beck, Yaffa , Nimrod, Abraham
IPC分类号： A61K38/44
CPC分类号： C12N9/0089 , A61K38/446
摘要： A double-stranded cDNA molecule which includes DNA encoding human manganese superoxide dismutase has been created. The sequence of one strand of a double-stranded DNA molecule which encodes human manganese superoxide dismutase has been discovered. Such molecules may be introduced in prokaryotic, e.g., bacterial, or eucaryotic, e.g., yeast or mammalian, cells and the resulting cells cultured or grown under suitable conditions so as to produce human manganese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants which may then be recovered. By this invention, human manganese superoxide dismutase gene fragments from various plasmids may be ligated to yield a complete genomic human manganese superoxide dismutase gene fragment. Human manganese superoxide dismutase, analogs, or mutants may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs, or treat inflammations. The invention also concerns a method of producing enzymatically active human manganese superoxide dismutase compounds in a bacterial cell which contains and is capable of expressing a DNA sequence encoding the superoxide dismutase compound by maintaining the bac terial cell under suitable conditions and in a suitable production medium. The production medium is supplemented with an amount of Mn⁺⁺ so that the concentration of Mn⁺⁺ in the medium is greater than about 2 ppm. Genomic manganese superoxide dismutase DNA should also be capable of expression in eucaryotic cells under suitable conditions. This invention also concerns a method of recovering purified enzymatically active manganese superoxide dismutase, analogs or mutants from bacterial cells. It should also be possible to recover manganese SOD from genomic manganese superoxide dismutase DNA expressed in eucaryotic cells using similar methods. The invention further concerns uses of the human manganese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants, various new analogs, compositions of the compounds and methods of treating various disorders using these compositions.
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56.发明公开
公开(公告)号：EP0259485A1
公开(公告)日：1988-03-16
申请号：EP87902255.0
申请日：1987-03-13
CPC分类号： A61K31/715 , A01N43/16 , C08B37/0072
摘要： Sels de métaux lourds de l'acide hyaluronique et notamment sels d'argent, d'or, de cérium et de tungstène de l'acide hyaluronique. Ces sels de métaux lourds de l'acide hyaluronique sont utiles en tant qu'agents antimicrobiens. L'hyaluronate d'or peut également être utilisé pour traiter l'arthrite. L'invention se rapporte également à des procédés de production de sels d'argent de l'acide hyaluronique ainsi qu'à des compositions contenant l'hyaluronate d'argent ou l'hyaluronate d'or. L'invention se rapporte également à des sels de métaux lourds possédant des moitiés d'hyaluronates marquées radioactivement.
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57.发明公开METHOD OF PRODUCING HIGH MOLECULAR WEIGHT SODIUM HYALURONATE BY FERMENTATION OF $i(STREPTOCOCCUS) 失效用于生产高分子量透明质酸钠BY链球菌的的外观。
公开(公告)号：EP0211037A1
公开(公告)日：1987-02-25
申请号：EP86900929.0
申请日：1986-01-16
IPC分类号： C12N15 , A61K8 , A61K31 , A61P27 , A61P29 , A61Q19 , C08B37 , C12N1 , C12P19 , C12Q1 , C12R1
CPC分类号： A61K31/715 , A61K8/735 , A61K2800/75 , A61Q19/00 , C08B37/0072 , C12P19/26 , C12R1/46 , Y10S435/803 , Y10S435/818 , Y10S435/885
摘要： Un nouveau microorganisme surtout, le Streptococcus zooepidemicus HA-116 ATCC 39920, produit de grandes quantités d'acide hyalyronique de poids moléculaire élevé. L'invention décrit un procédé permettant d'obtenir ces microorganismes, ainsi qu'un procédé de production d'hyaluronate de sodium consistant à cultiver sous agitation vigoureuse un microorganisme du genre Streptococcus dans des conditions et dans un milieu nutritif appropriés contenant un composant de sucre comme source de carbone. Le composant du sucre a une teneur sensiblement constante comprise entre 0,2 et 10 grammes par litre. Le milieu présente un pH sensiblement constant compris entre 6,0 et 7,5 et comprend une teneur en ions magnésium sensiblement constante et supérieure à 0,05 grammes par litre. L'hyaluronate de sodium excrété dans le milieu par l'organisme est purifié par des procédés consistant à précipiter, redissoudre et reprécipiter l'hyaluronate. On obtient ainsi des compositions à base d'hyaluronate de sodium caractérisées par l'absence de pyrogénicité et n'irritant pas la peau.
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