公开(公告)号：US5198361A

公开(公告)日：1993-03-30

申请号：US635413

申请日：1991-01-03

申请人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

发明人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

IPC分类号： A61K38/00 ,  C07K14/61 ,  C12N15/73

CPC分类号： C07K14/61 ,  C12N15/73 ,  A61K38/00

摘要： An improved vector upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of a desired gene inserted into the vector and production of polypeptide encoded by the gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: a DNA sequence which contains the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the desired gene capable of binding to ribosomes within the host cell; an ATG initiation codon or a DNA sequence which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a restriction enzyme site for inserting the desired gene into the vector in phase with the ATG initiation codon; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the vector is present in the host cell.

摘要翻译： 在引入含有不耐热阻遏物CI的合适的细菌宿主中时，改进的载体使宿主细胞在将宿主细胞的温度升高至阻遏物被破坏的温度时能够实现插入到载体中的所需基因的表达 并产生由该基因编码的多肽。 载体是双链DNA分子，其以5'至3'的顺序包括以下：含有来自λ噬菌体的启动子和操纵子PLOL的DNA序列; 用于结合由宿主细胞产生的抗终止子N蛋白的N利用位点; 一个DNA序列，其含有一个核糖体结合位点，用于使所需基因的mRNA能够结合宿主细胞内的核糖体; ATG起始密码子或将所需基因插入载体后转化为ATG起始密码子的DNA序列; 用于将所需基因插入与ATG起始密码子同相载体的限制酶位点; 另外还包含含有能够在宿主细胞中自主复制的细菌质粒的复制起点的DNA序列和含有与可选择或可识别的性状相关的基因的DNA序列，所述基因在载体存在于宿主细胞中时显示 。

公开(公告)号：US4871835A

公开(公告)日：1989-10-03

申请号：US691230

申请日：1985-01-14

申请人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Pinhas E. Zeelon ,  Menachem Zeevi

发明人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Pinhas E. Zeelon ,  Menachem Zeevi

IPC分类号： A61K38/00 ,  C07K14/61 ,  C12N15/73 ,  C12P21/02

CPC分类号： C12P21/02 ,  C07K14/61 ,  C12N15/73 ,  A61K38/00 ,  Y10S530/806 ,  Y10S530/808 ,  Y10S530/82 ,  Y10S930/12

摘要： Analogs of hGH having the activity of naturally occurring hGH and a similar amino acid sequence varying from the sequence of natural hGh by the addition of one or more amino acids, e.g. methione or methionine-leucine, to the N-terminus of natural hGH have been produced, recovered and purified. Such analogs may be incorporated into pharmaceutical compositions and administered to a subject to increase the level of hGH in the subject.Analogs of hGH which comprise the amino acid sequence of natural hGH from the N-terminus of which one or more amino acids have been deleted, e.g. Met.sup.14 hGh, have been produced, recovered and purified. Such analogs may be incorporated into pharmaceutical compositions and administered to a subject to lower the level of hGH in the subject.A plasmid has been constructed which directs the expression of an analog of hGH having the amino acid sequence methionine-leucine added to the N-terminus of natural hGH. This plasmid has been introduced into Escherichia coli, the resulting Escherichia coli grown and the analog recovered and purified.

摘要翻译： 具有天然存在的hGH活性的hGH的类似物和通过加入一种或多种氨基酸例如天然hGh的序列而变化的类似氨基酸序列，例如， 甲硫氨酸或甲硫氨酸 - 亮氨酸已经产生，回收和纯化到天然hGH的N末端。 这样的类似物可以并入药物组合物中并且施用于受试者以增加受试者中hGH的水平。 包含其N末端的天然hGH的氨基酸序列的类似物，其一个或多个氨基酸已被缺失，例如， Met14hGh已经生产，回收和纯化。 这样的类似物可以并入药物组合物中并施用于受试者以降低受试者的hGH水平。 已经构建了一种质粒，其将氨基酸序列甲硫氨酸 - 亮氨酸类似物的表达指导到天然hGH的N末端。 将该质粒导入大肠杆菌中，生成大肠杆菌，回收并纯化。

公开(公告)号：US4831120A

公开(公告)日：1989-05-16

申请号：US752441

申请日：1985-07-03

申请人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

发明人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

IPC分类号： C12N15/09 ,  A61K35/74 ,  A61K38/00 ,  A61K38/27 ,  C07H21/04 ,  C07K14/00 ,  C07K14/485 ,  C07K14/52 ,  C07K14/575 ,  C07K14/61 ,  C12N1/20 ,  C12N1/21 ,  C12N9/02 ,  C12N15/00 ,  C12N15/53 ,  C12N15/73 ,  C12P21/02 ,  C12R1/19 ,  C12R1/91

CPC分类号： C12N9/0089 ,  C07K14/61 ,  C12N15/73 ,  A61K38/00 ,  Y10S530/82 ,  Y10S530/825 ,  Y10S930/12

摘要： An improved vector upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of a desired gene inserted into the vector and production of polypeptide encoded by the gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: a DNA sequence which contains the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the desired gene capable of binding to ribosomes within the host cell; an ATG initiation codon or a DNA sequence which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a restriction enzyme site for inserting the desired gene into the vector in phase with the ATG initiation codon; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the vector is present in the host cell.

摘要翻译： 在引入含有不耐热阻遏物CI的合适的细菌宿主中时，改进的载体使宿主细胞在将宿主细胞的温度升高至阻遏物被破坏的温度时能够实现插入到载体中的所需基因的表达 并产生由该基因编码的多肽。 载体是双链DNA分子，其以5'至3'的顺序包括以下：含有来自λ噬菌体的启动子和操纵子PLOL的DNA序列; 用于结合由宿主细胞产生的抗终止子N蛋白的N利用位点; 一个DNA序列，其含有一个核糖体结合位点，用于使所需基因的mRNA能够结合宿主细胞内的核糖体; ATG起始密码子或将所需基因插入载体后转化为ATG起始密码子的DNA序列; 用于将所需基因插入与ATG起始密码子同相载体的限制酶位点; 另外还包含含有能够在宿主细胞中自主复制的细菌质粒的复制起点的DNA序列和含有与可选择或可识别的性状相关的基因的DNA序列，所述基因在载体存在于宿主细胞中时显示 。

公开(公告)号：US4997916A

公开(公告)日：1991-03-05

申请号：US320320

申请日：1989-03-07

申请人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

发明人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

IPC分类号： A61K38/00 ,  C07K14/61 ,  C12N15/73

CPC分类号： C07K14/61 ,  C12N15/73 ,  A61K38/00 ,  Y10S530/82 ,  Y10S530/825

摘要： A method is provided for recovering a purified animal growth hormone or a polypeptide analog thereof having substantially the same amino acid sequence as, and the biological activity of, the corresponding naturally-occurring animal growth hormone from a bacterial cell in which the animal growth hormone or polypeptide analog has been produced by means of expression of a plasmid encoding the hormone or polypeptide analog which comprises: (a) disrupting the cell wall of the bacterial cell in a buffered neutral pH solution so as to produce a lysate containing precipitated hormone or polypeptide analog; (b) recovering the resulting precipitated hormone or polypeptide analog; (c) suspending the precipitated hormone or polypeptide analog so recovered in distilled water; (d) treating the resulting precipitate-containing suspension with a sodium hydroxide solution having an alkaline pH of about 11.8 so as to solubilize the precipitate and thus the hormone or polypeptide analog contained therein; (e) separating the solubilized hormone or polypeptide analog from other soluble components by gel filtration chromatography; and (f) subjecting the hormone or polypeptide analog thus separated to ion exchange chromatography to purify the hormone or analog and thereby recover purified hormone or polypeptide analog.

摘要翻译： 提供了一种用于从动物生长激素或细菌细胞的细菌细胞中回收纯化的动物生长激素或其多肽类似物，其具有与相应天然存在的动物生长激素基本上相同的氨基酸序列和生物学活性， 已经通过表达编码激素或多肽类似物的质粒产生多肽类似物，其包括：（a）在缓冲的中性pH溶液中破坏细菌细胞的细胞壁，以产生含有沉淀激素或多肽类似物的裂解物 ; （b）回收所得沉淀的激素或多肽类似物; （c）将沉淀的激素或多肽类似物悬浮于蒸馏水中; （d）用碱性pH为约11.8的氢氧化钠溶液处理所得沉淀物的悬浮液，以溶解沉淀物，从而溶解其中所含的激素或多肽类似物; （e）通过凝胶过滤色谱法将溶解的激素或多肽类似物与其他可溶组分分离; 和（f）使由此分离的激素或多肽类似物经离子交换色谱纯化激素或类似物，从而回收纯化的激素或多肽类似物。

公开(公告)号：US5256546A

公开(公告)日：1993-10-26

申请号：US840579

申请日：1992-02-24

申请人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

发明人： Haim Aviv ,  Marian Gorecki ,  Avigdor Levanon ,  Amos Oppenheim ,  Tikva Vogel ,  Elisha Zeelon ,  Menachem Zeevi

IPC分类号： A61K38/00 ,  C07K14/61 ,  C12N15/18 ,  C12N15/73 ,  C12P21/02 ,  C12N15/03

CPC分类号： C12N15/73 ,  C07K14/61 ,  A61K38/00

摘要： A plasmid for the production of porcine growth hormone or an analog thereof which upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of DNA encoding porcine growth hormone and production of porcine growth hormone or analogs thereof. The plasmid is a double-stranded DNA molecule which includes in 5' to 3' the following: a DNA sequence which contains the promoter and operator PLOL from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the gene encoding porcine growth hormone capable of binding to ribosomes within the host cell; an ATG initiation codon; a restriction enzyme site for inserting the gene encoding porcine growth hormone into the plasmid in phase with the ATG initiation codon; a gene encoding porcine growth hormone; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the plasmid is present in the host cell.

摘要翻译： 用于生产猪生长激素或其类似物的质粒，其在引入含有不耐热阻遏物CI的合适的细菌宿主中时，使宿主细胞能够在将宿主细胞的温度升高至阻遏物被破坏的温度时， 实现编码猪生长激素的DNA的表达和猪生长激素或其类似物的产生。 质粒是双链DNA分子，其包括以下5'至3'：含有来自λ噬菌体的启动子和操纵子PLOL的DNA序列; 用于结合由宿主细胞产生的抗终止子N蛋白的N利用位点; 包含用于使编码能够结合宿主细胞内核糖体的猪生长激素的基因的mRNA的核糖体结合位点的DNA序列; ATG起始密码子; 用于将ATG起始密码子与编码猪生长激素的基因插入质粒的限制酶切位点; 一种编码猪生长激素的基因; 另外还包含一个DNA序列，其含有能够在宿主细胞中自主复制的细菌质粒的复制起点，以及含有与选择性或可识别性状相关的基因的DNA序列，当该质粒存在于宿主细胞中时 。