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公开(公告)号：US5583010A
公开(公告)日：1996-12-10
申请号：US73799
申请日：1993-06-10
申请人： William R. Baumbach , John R. Zysk , Michael O. Thorner , Bruce D. Gaylinn , Kevin R. Lynch , Jeffrey K. Harrison
发明人： William R. Baumbach , John R. Zysk , Michael O. Thorner , Bruce D. Gaylinn , Kevin R. Lynch , Jeffrey K. Harrison
CPC分类号： C07K14/723 , C07K14/60 , G01N33/74 , A61K38/00 , G01N2333/60 , G01N2333/726
摘要： Growth hormone releasing hormone (GHRH) receptor binding has been characterized using a unique binding assay utilizing iodinated GHRH probes. Photoaffinity GHRH probes have been constructed which allow for photolabeling and characterization of the receptor. In addition, high affinity biotinylated GHRH analogs have been constructed. Solubilization of GHRH-R/GHRH complexes and extraction of specifically bound GHRH using a mild detergent solution, followed by affinity chromotography, leads to a substantially purified GHRH-R isolate. Electrophoretic treatment of the GHRH-R isolate produces GHRH-R of sufficient purity to conduct sequencing of the receptor. Cloning of a gene encoding for polypeptides (protein or fragments thereof) having GHRH-R activity is accomplished using a bacterial host, and the cloned gene is expressed in a mammalian cell line.
摘要翻译： 生长激素释放激素（GHRH）受体结合已经通过使用碘化GHRH探针的独特结合测定来表征。 已经构建了能够对受体进行光贴标和表征的光亲和性GHRH探针。 此外，已经构建了高亲和力的生物素化GHRH类似物。 GHRH-R / GHRH复合物的溶解和使用温和洗涤剂溶液提取特异性结合的GHRH，然后进行亲和层析，得到基本纯化的GHRH-R分离物。 GHRH-R分离物的电泳处理产生足够纯度的GHRH-R以进行受体的测序。 使用细菌宿主完成编码具有GHRH-R活性的多肽（蛋白质或其片段）的基因的克隆，并且克隆的基因在哺乳动物细胞系中表达。
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公开(公告)号：US5612470A
公开(公告)日：1997-03-18
申请号：US947672
申请日：1992-09-18
IPC分类号： G01N33/53 , A61K38/00 , C07K1/113 , C07K1/22 , C07K14/575 , C07K14/60 , C07K14/61 , C07K14/655 , C07K14/705 , C07K14/72 , C07K17/02 , C12N15/12 , G01N33/52 , G01N33/74
CPC分类号： C07K14/60 , C07K14/723 , G01N33/74 , A61K38/00 , G01N2333/60 , G01N2333/726
摘要： Growth hormone releasing hormone (GHRH) receptor binding has been characterized using a unique binding assay utilizing iodinated GHRH probes. Photoaffinity GHRH probes have been constructed which allow for photolabeling and characterization of the receptor. In addition, high affinity biotinylated GHRH analogs have been constructed. Solubilization of GHRH-R/GHRH complexes and extraction of specifically bound GHRH using a mild detergent solution, followed by affinity chromotography, leads to a substantially purified GHRH-R isolate. Electrophoretic treatment of the GHRH-R isolate produces GHRH-R of sufficient purity to conduct sequencing of the receptor. Cloning of a gene encoding for polypeptides (protein or fragments thereof) having GHRH-R activity is accomplished using a bacterial host, and the cloned gene is expressed in a mammalian cell line.
摘要翻译： 生长激素释放激素（GHRH）受体结合已经通过使用碘化GHRH探针的独特结合测定来表征。 已经构建了能够对受体进行光贴标和表征的光亲和性GHRH探针。 此外，已经构建了高亲和力的生物素化GHRH类似物。 GHRH-R / GHRH复合物的溶解和使用温和洗涤剂溶液提取特异性结合的GHRH，随后进行亲和层析，得到基本纯化的GHRH-R分离物。 GHRH-R分离物的电泳处理产生足够纯度的GHRH-R以进行受体的测序。 使用细菌宿主完成编码具有GHRH-R活性的多肽（蛋白质或其片段）的基因的克隆，并且克隆的基因在哺乳动物细胞系中表达。
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公开(公告)号：US5591641A
公开(公告)日：1997-01-07
申请号：US946232
申请日：1992-09-15
IPC分类号： G01N33/53 , A61K38/00 , A61K38/25 , C07K1/113 , C07K1/22 , C07K14/575 , C07K14/60 , C07K14/61 , C07K14/655 , C07K14/705 , C07K14/72 , C07K17/02 , C12P21/02 , G01N33/52 , G01N33/74 , C12N15/12
CPC分类号： C07K14/60 , C07K14/723 , G01N33/74 , A61K38/00 , G01N2333/60 , G01N2333/726
摘要： Growth hormone releasing hormone (GHRH) receptor binding has been characterized using a unique binding assay utilizing iodinated GHRH probes. Photoaffinity GHRH probes have been constructed which allow for photolabeling and characterization of the receptor. In addition, high affinity biotinylated GHRH analogs have been constructed. Solubilization of GHRH-R/GHRH complexes and extraction of specifically bound GHRH using a mild detergent solution, followed by affinity chromotography, leads to a substantially purified GHRH-R isolate. Electrophoretic treatment of the GHRH-R isolate produces GHRH-R of sufficient purity to conduct sequencing of the receptor. Cloning of a gene encoding for polypeptides (protein or fragments thereof) having GHRH-R activity is accomplished using a bacterial host, and the cloned gene is expressed in a mammalian cell line.
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4.发明授权
公开(公告)号：US5644046A
公开(公告)日：1997-07-01
申请号：US432043
申请日：1995-05-01
申请人： Michael Oliver Thorner , Bruce David Gaylinn , Jeffrey K. Harrison , Kevin R. Lynch , John Ronald Zysk
发明人： Michael Oliver Thorner , Bruce David Gaylinn , Jeffrey K. Harrison , Kevin R. Lynch , John Ronald Zysk
IPC分类号： G01N33/53 , A61K38/00 , C07K1/113 , C07K1/22 , C07K14/575 , C07K14/60 , C07K14/61 , C07K14/655 , C07K14/705 , C07K14/72 , C07K17/02 , C12N15/12 , G01N33/52 , G01N33/74
CPC分类号： C07K14/60 , C07K14/723 , G01N33/74 , A61K38/00 , G01N2333/60 , G01N2333/726
摘要： Growth hormone releasing hormone (GHRH) receptor binding has been characterized using a unique binding assay utilizing iodinated GHRH probes. Photoaffinity GHRH probes have been constructed which allow for photolabeling and characterization of the receptor. In addition, high affinity biotinylated GHRH analogs have been constructed. Solubilization of GHRH-R/GHRH complexes and extraction of specifically bound GHRH using a mild detergent solution, followed by affinity chromatography, leads to a substantially purified GHRH-R isolate. Electrophoretic treatment of the GHRH-R isolate produces GHRH-R of sufficient purity to conduct sequencing of the receptor. Cloning of a gene encoding for polypeptides (protein or fragments thereof) having GHRH-R activity is accomplished using a bacterial host, and the cloned gene is expressed in a mammalian cell line.
摘要翻译： 生长激素释放激素（GHRH）受体结合已经通过使用碘化GHRH探针的独特结合测定来表征。 已经构建了能够对受体进行光贴标和表征的光亲和性GHRH探针。 此外，已经构建了高亲和力的生物素化GHRH类似物。 GHRH-R / GHRH复合物的溶解和使用温和洗涤剂溶液提取特异性结合的GHRH，然后进行亲和层析，得到基本纯化的GHRH-R分离物。 GHRH-R分离物的电泳处理产生足够纯度的GHRH-R以进行受体的测序。 使用细菌宿主完成编码具有GHRH-R活性的多肽（蛋白质或其片段）的基因的克隆，并且克隆的基因在哺乳动物细胞系中表达。
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