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    Recombinant human granulocyte-macrophage-colony stimulating factor (GM-CSF)
    11.
    发明授权
    Recombinant human granulocyte-macrophage-colony stimulating factor (GM-CSF) 失效
    重组人粒细胞巨噬细胞集落刺激因子（GM-CSF）

    公开(公告)号：US5891429A

    公开(公告)日：1999-04-06

    申请号：US466308

    申请日：1995-06-06

    申请人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    发明人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    IPC分类号： A61K38/00 ,  C07K14/535 ,  C12N15/27 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  A61K38/19 ,  C07K1/20

    CPC分类号： C12N15/85 ,  C07K14/535 ,  C12N15/70 ,  C12N15/81 ,  A61K38/00 ,  C12N2800/206 ,  C12N2830/00 ,  C12N2830/702 ,  C12N2840/105 ,  C12N2840/44

    摘要： A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises:preparing RNA from a cell that produces CSF;preparing polyadenylated messenger RNA from said RNA;preparing single stranded cDNA from said messenger RNA;converting the single stranded cDNA to double stranded cDNA;inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies;picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool;transfecting the plasmid DNA into suitable host cells for expressing CSF protein;culturing the transfected cells and assaying the supernatant for CSF activity; andselecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line. The invention also provides a method of purifying the CSF proteins and the purified proteins so produced.

    摘要翻译： 描述了制备和分离含有CSF / cDNA的转化载体的方法。 该方法包括：从产生CSF的细胞制备RNA; 从所述RNA制备聚腺苷酸化的信使RNA; 从所述信使RNA制备单链cDNA; 将单链cDNA转化为双链cDNA; 将双链cDNA插入转化载体并用所述载体转化细菌以形成菌落; 分别取200至500个菌落的池，每个池中分离出质粒DNA; 将质粒DNA转染到合适的宿主细胞中以表达CSF蛋白; 培养转染的细胞并测定上清液的CSF活性; 并选择CSF阳性池并筛选用于制备池的菌落以鉴定具有CSF活性的菌落。 还描述了编码具有CSF活性的蛋白质（即CSF / cDNA）的cDNA，用含有这种CSF / cDNA的重组载体转化的微生物或细胞系，以及通过培养表达所述CSF / cDNA来产生CSF蛋白的方法 微生物或细胞系。 本发明还提供了纯化CSF蛋白质和如此制备的纯化蛋白质的方法。

    Recombinant primate granulocyte macrophage-colony stimulating factor
    14.
    发明授权
    Recombinant primate granulocyte macrophage-colony stimulating factor 失效
    重组灵长类动物粒细胞巨噬细胞集落刺激因子

    公开(公告)号：US5942221A

    公开(公告)日：1999-08-24

    申请号：US468694

    申请日：1995-06-06

    申请人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong

    发明人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong

    IPC分类号： C12P21/02 ,  A61K38/00 ,  A61K38/17 ,  C07K14/535 ,  C12N15/27 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  A61K38/19

    CPC分类号： C07K14/535 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  A61K38/00 ,  C12N2830/42 ,  C12N2840/105 ,  Y10S514/885

    摘要： A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises: preparing RNA from a cell that produces CSF; preparing polyadenylated messenger RNA from said RNA; preparing single stranded cDNA from said messenger RNA; converting the single stranded cDNA to double stranded cDNA; inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies; picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool; transfecting the plasmid DNA into suitable host cells for expressing CSF protein; culturing the transfected cells and assaying the supernatant for CSF activity; and selecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line.

    摘要翻译： 描述了制备和分离含有CSF / cDNA的转化载体的方法。 该方法包括：从产生CSF的细胞制备RNA; 从所述RNA制备聚腺苷酸化的信使RNA; 从所述信使RNA制备单链cDNA; 将单链cDNA转化为双链cDNA; 将双链cDNA插入转化载体并用所述载体转化细菌以形成菌落; 分别取200至500个菌落的池，每个池中分离出质粒DNA; 将质粒DNA转染到合适的宿主细胞中以表达CSF蛋白; 培养转染的细胞并测定上清液的CSF活性; 并选择CSF阳性池并筛选用于制备池的菌落以鉴定具有CSF活性的菌落。 还描述了编码具有CSF活性的蛋白质（即CSF / cDNA）的cDNA，用含有这种CSF / cDNA的重组载体转化的微生物或细胞系，以及通过培养表达所述CSF / cDNA来产生CSF蛋白的方法 微生物或细胞系。

    DNA encoding GM-CSF and a method of producing GM-CSF protein
    17.
    发明授权
    DNA encoding GM-CSF and a method of producing GM-CSF protein 失效
    编码GM-CSF的DNA和生产GM-CSF蛋白的方法

    公开(公告)号：US5908763A

    公开(公告)日：1999-06-01

    申请号：US287019

    申请日：1994-08-08

    申请人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    发明人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    IPC分类号： A61K38/00 ,  C07K14/535 ,  C12N15/27 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  C07K14/53 ,  C12N15/63

    CPC分类号： C12N15/85 ,  C07K14/535 ,  C12N15/70 ,  C12N15/81 ,  A61K38/00 ,  C12N2800/206 ,  C12N2830/00 ,  C12N2830/702 ,  C12N2840/105 ,  C12N2840/44

    摘要： A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises:preparing RNA from a cell that produces CSF;preparing polyadenylated messenger RNA from said RNA;preparing single stranded cDNA from said messenger RNA;converting the single stranded cDNA to double stranded cDNA;inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies;picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool;transfecting the plasmid DNA into suitable host cells for expressing CSF protein;culturing the transfected cells and assaying the supernatant for CSF activity; andselecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line. The invention also provides a method of purifying the CSF proteins and the purified proteins so produced.

    摘要翻译： 描述了制备和分离含有CSF / cDNA的转化载体的方法。 该方法包括：从产生CSF的细胞制备RNA; 从所述RNA制备聚腺苷酸化的信使RNA; 从所述信使RNA制备单链cDNA; 将单链cDNA转化为双链cDNA; 将双链cDNA插入转化载体并用所述载体转化细菌以形成菌落; 分别取200至500个菌落的池，每个池中分离出质粒DNA; 将质粒DNA转染到合适的宿主细胞中以表达CSF蛋白; 培养转染的细胞并测定上清液的CSF活性; 并选择CSF阳性池并筛选用于制备池的菌落以鉴定具有CSF活性的菌落。 还描述了编码具有CSF活性的蛋白质（即CSF / cDNA）的cDNA，用含有这种CSF / cDNA的重组载体转化的微生物或细胞系，以及通过培养表达所述CSF / cDNA来产生CSF蛋白的方法 微生物或细胞系。 本发明还提供了纯化CSF蛋白质和如此制备的纯化蛋白质的方法。