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    DNA encoding human interleukin 1 alpha and the amino acid chain corresponding thereto and vectors and hosts containing such DNA; and the preparatiion thereof
    2.
    发明公开
    DNA encoding human interleukin 1 alpha and the amino acid chain corresponding thereto and vectors and hosts containing such DNA; and the preparatiion thereof 失效
    用于α-编码人白细胞介素1的DNA和其相应的氨基酸链，含有这种DNA的载体和宿主以及它们的制备。

    公开(公告)号：EP0188864A1

    公开(公告)日：1986-07-30

    申请号：EP85303701.8

    申请日：1985-05-24

    申请人： IMMUNEX CORPORATION

    发明人： Cerretti, Douglas P. ,  Conlon III, Paul J. ,  Cosman, David J. ,  Grabstein, Kenneth H. ,  Hopp, Thomas P. ,  Kronheim, Shirley R. ,  Larsen, Alf D. ,  March, Carl J. ,  Mosley, Bruce A. ,  Price, Virginia L.

    IPC分类号： C12N15/25 ,  C07H21/04 ,  C12P21/02 ,  C07K13/00

    CPC分类号： C07K14/545

    摘要： Double-stranded cDNA is prepared from polyadenylated RNA extracted from activated human peripheral blood adherent mononuclear cells. The cDNA is inserted within a plasmid vector and then the recombinant plasmid employed to transform an appropriate host. Transformed hosts are identified and grouped into pools. Plasmid DNA prepared from these pools is hybridized with total mRNA from human peripheral blood cells. Pools of host cells that provide a positive signal from the hybrid section method are subdivided and the procedure repeated until a single positive colony is isolated. Plasmid DNA is prepared from this colony and radiolabeled for use as a probe to rescreen the cDNA library for a larger clone. Also a radiolabeled, synthetic oligonucleotide probe corresponding to a portion of the initially isolated clone is used to further screen the cDNA library for additional clones. Portions of the additional clones are then employed as labeled probes to further screen the cDNA library. By this process, a single clone spanning the entire open reading frame of the IL-1 gene was isolated as well as several other clones composing a portion of the gene. The entire open reading frame and the coding region of the IL-1 gene was inserted into expression vectors for expression of functional IL-1.

    摘要翻译： 双链cDNA是从激活的人外周血单核细胞贴壁提取聚腺苷酸化RNA制备。 将cDNA插入一个质粒载体中，然后用的重组质粒转化到合适的宿主。 转化宿主被识别并分组到池。 从池合成制备质粒DNA杂交从人外周血细胞总mRNA。 宿主细胞的池确实提供来自混合部分方法的阳性信号细分并且重复该程序，直到单个阳性菌落分离。 质粒DNA从该菌落并制备放射标记用作探针来rescreen cDNA文库获得较大克隆。 这样的放射性标记的，合成的寡核苷酸探针对应于最初分离克隆的一部分用于进一步筛选cDNA文库为额外的克隆。 然后将附加的克隆的部分被用作标记的探针つweiterer筛选cD​​NA文库。 通过这种方法，单个克隆跨越其分离，以及构成该基因的一部分，其他的几个克隆的IL-1基因的整个开放阅读框。 整个开放读框和其插入表达载体中用于功能性IL-1的表达的IL-1基因的编码区。

    Purified interleukin 1 and DNA coding for interleukin 1 and their preparation, vectors containing such DNA and their preparation and use in transforming hosts to permit expression of interleukin 1
    4.
    发明公开
    Purified interleukin 1 and DNA coding for interleukin 1 and their preparation, vectors containing such DNA and their preparation and use in transforming hosts to permit expression of interleukin 1 失效
    纯化的白细胞介素-1和白介素-1编码DNA和其制造，例如含有DNA的载体和它们的生产和使用用于转化宿主，其允许白介素-1的表达。

    公开(公告)号：EP0456332A1

    公开(公告)日：1991-11-13

    申请号：EP91201769.6

    申请日：1985-03-13

    申请人： IMMUNEX CORPORATION

    发明人： Cerretti, Douglas P. ,  Conlon III, Paul J. ,  Cosman, David J. ,  Grabstein, Kenneth H. ,  Hopp, Thomas P. ,  Kronheim, Shirley R. ,  Larsen, Alf D. ,  March, Carl J. ,  Price, Virgina L.

    IPC分类号： C12N15/00 ,  C12P21/02 ,  C07K13/00 ,  C12N1/00

    CPC分类号： C07K14/545 ,  C12N15/81

    摘要： Interleukin 1 is purified to homogeneity by use of various techniques including ion exchange chromatography and dye-ligand affinity chromatography, thereby enabling the amino acid composition of this protein to be ascertained and its amino acid sequence to be partially determined. Double-stranded cDNA is prepared from polyadenylated RNA extracted from activated human peripheral blood adherent mononuclear cells. The cDNA is inserted within a plasmid vector and then the recombinant plasmid employed to transform an appropriate host. Plasmid DNA prepared from pools of transformed hosts is hybridized with a labeled, synthetic oligonucleotide probe corresponding to a portion of the ascertained amino acid sequence of the interleukin 1 protein. Pools of host cells that provide a positive signal to the probe are identified, plated out and then employed in direct bacterial colony hybridization with the same probe, thereby to isolate the particular positive colony. Plasmid DNA is prepared from this colony and characterized by restriction enzyme mapping and sequenced. The coding region for the IL-1 gene is inserted into a shuttle vector for amplification of the vector followed by expression of functional IL-1.

    摘要翻译： 白细胞介素1通过使用包括离子交换色谱法和染料配体亲和色谱法的各种技术纯化至均一，从而使该蛋白质待确定和其氨基酸序列的氨基酸组合物是部分确定性开采。 双链cDNA是从激活的人外周血单核细胞贴壁提取聚腺苷酸化RNA制备。 将cDNA插入一个质粒载体中，然后用的重组质粒转化到合适的宿主。 从转化的宿主池制备质粒DNA杂交，用标记的，合成的寡核苷酸探针对应于白介素1蛋白质的确定氨基酸序列的一部分。 宿主细胞的池确实提供正信号到探针被识别，镀测试出，然后在直接细菌菌落杂交采用具有相同的，由此以分离的特定阳性菌落。 质粒DNA从该集落制备并通过限制性酶图谱并测序特征的。 用于IL-1基因的编码区插入到穿梭载体用于随后功能性IL-1的表达载体的扩增。

    Interleukin-2 receptor and DNA encoded therefor and their preparation
    7.
    发明公开
    Interleukin-2 receptor and DNA encoded therefor and their preparation 失效
    白细胞介素-2受体激动剂，dafürkodierende DNS und deren Herstellung。

    公开(公告)号：EP0162699A2

    公开(公告)日：1985-11-27

    申请号：EP85303575.6

    申请日：1985-05-21

    申请人： IMMUNEX CORPORATION

    发明人： Cerretti, Douglas P. ,  Cosman, David J. ,  Dower, Steven K. ,  March, Carl J. ,  Urdal, David L. ,  Larsen, Alf D.

    IPC分类号： C12N15/12 ,  C12N15/85 ,  C12N5/10 ,  C12P21/02

    CPC分类号： C07K14/7155 ,  C07K16/2866

    摘要： Interleukin-2 receptor derived from normal and malignant cells has been purified by use of various techniques including affinity chromatography in conjunction with a monoclonal antibody directed to the receptor. The purification process also includes reversed phased high performance liquid chromatography. By these techniques, interleukin-2 receptor has been purified to homogeneity. The high purification of the interleukin-2 receptor has made possible the sequencing of the amino acid residues at the N-terminal of this protein molecule. Double-stranded cDNA is prepared from polyadenylated RNA extracted from cell lines or other sources known to produce IL-2 receptor. The cDNA is inserted within a plasmid vector and then the recombinant plasmid employed to transform an appropriate host. Transformed hosts are identified and grouped into pools. Plasmid DNA prepared from these pools is hybridized with a labeled synthetic oligonucleotide probe corresponding to a portion of the amino acid sequence of the purified IL-2 receptor. The cDNA clone isolated with the probe is characterized by restriction enzyme mapping and sequenced by chain-termination method. The particular DNA clone that actually contains the gene coding for the functional IL-2 receptor is identified by expressing the clones in COS-7 monkey kidney cells and assaying for the expressed IL-2 receptor by its ability to bind IL-2 or a monoclonal antibody directed against the IL-2 receptor.

    摘要翻译： 来自正常和恶性细胞的白细胞介素-2受体已经通过使用各种技术进行纯化，包括亲和层析与针对受体的单克隆抗体。 纯化方法还包括反相高效液相色谱法。 通过这些技术，白细胞介素-2受体已经被纯化到同质性。 白细胞介素-2受体的高度纯化使得可以对该蛋白质分子的N-末端的氨基酸残基进行测序。 从从已知产生IL-2受体的细胞系或其他来源提取的聚腺苷酸化RNA制备双链cDNA。 将cDNA插入质粒载体中，然后将重组质粒用于转化合适的宿主。 已识别转换的主机并将其分组到池中。 从这些池制备的质粒DNA与对应于纯化的IL-2受体的氨基酸序列的一部分的标记的合成寡核苷酸探针杂交。 用探针分离的cDNA克隆的特征在于限制酶作图并通过链终止法进行测序。 通过在COS-7猴肾细胞中表达克隆并通过其结合IL-2或单克隆抗体的能力测定表达的IL-2受体来鉴定实际含有编码功能性IL-2受体的基因的特定DNA克隆 针对IL-2受体的抗体。