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1.
公开(公告)号:US06864060B1
公开(公告)日:2005-03-08
申请号:US08946298
申请日:1997-10-07
申请人: Dana M. Fowlkes , James R. Broach , John P. Manfredi , Jeremy I. Paul , Joshua Trueheart , Christine A. Klein , Andrew J. M. Murphy
发明人: Dana M. Fowlkes , James R. Broach , John P. Manfredi , Jeremy I. Paul , Joshua Trueheart , Christine A. Klein , Andrew J. M. Murphy
IPC分类号: C07K14/395 , C07K14/47 , C12N15/81 , G01N33/50 , G01N33/569 , G12N15/81
CPC分类号: G01N33/50 , C07K14/395 , C07K14/4722 , C07K14/705 , C07K14/723 , C07K2319/00 , C07K2319/02 , C12N1/16 , C12N15/81 , C40B30/06 , G01N33/566 , G01N2333/726 , G01N2500/02
摘要: The present invention pertains to novel yeast cells which are useful for the expression of heterologous G protein coupled receptors. The yeast cells of the present invention can be used in screening assays which can be used to screen for modulators of G protein coupled receptors. Specifically, the invention provides novel yeast cells which express a heterologous G protein coupled receptor and mutant and/or chimeric G protein subunit molecules which serve to functionally integrate the heterologous into the pheromone signaling pathway of the yeast cell. The invention also provides for the expression of heterologous G protein coupled receptors which are functionally integrated into the yeast cell membrane using a yeast α factor leader sequence. Drug discovery assays using the subject yeast cells are also provided.
摘要翻译: 本发明涉及可用于表达异源G蛋白偶联受体的新型酵母细胞。 本发明的酵母细胞可用于筛选测定,其可用于筛选G蛋白偶联受体的调节剂。 具体地,本发明提供了表达异源G蛋白偶联受体和突变体和/或嵌合G蛋白亚基分子的新型酵母细胞,其用于将异源整合到酵母细胞的信息素信号通路中。 本发明还提供使用酵母α因子前导序列在功能上整合到酵母细胞膜中的异源G蛋白偶联受体的表达。 还提供使用本发明酵母细胞的药物发现测定。
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公开(公告)号:US07105309B2
公开(公告)日:2006-09-12
申请号:US10263341
申请日:2002-10-01
申请人: Dana M. Fowlkes , James R. Broach , John P. Manfredi , Christine A. Klein , Andrew J. M. Murphy , Jeremy I. Paul , Joshua Trueheart
发明人: Dana M. Fowlkes , James R. Broach , John P. Manfredi , Christine A. Klein , Andrew J. M. Murphy , Jeremy I. Paul , Joshua Trueheart
IPC分类号: G01N33/567
CPC分类号: C12N15/81 , C07K14/395 , C07K2319/02
摘要: Yeast cells are engineered to express both a surrogate of a pheromone system protein (e.g., enzymes involved in maturation of α-factor, transporters of a-factor, pheromone receptors, etc.) and a potential peptide modulator of the surrogate, in such a manner that the inhibition or activation of the surrogate affects a screenable or selectable trait of the yeast cells. Various additional features improve the signal-to-noise ratio of the screening/selection system.
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3.
公开(公告)号:US07361498B2
公开(公告)日:2008-04-22
申请号:US10967087
申请日:2004-10-15
申请人: Dana M. Fowlkes , James R. Broach , John P. Manfredi , Jeremy I. Paul , Joshua Trueheart , Christine A. Klein , Andrew J. M. Murphy
发明人: Dana M. Fowlkes , James R. Broach , John P. Manfredi , Jeremy I. Paul , Joshua Trueheart , Christine A. Klein , Andrew J. M. Murphy
CPC分类号: G01N33/50 , C07K14/395 , C07K14/4722 , C07K14/705 , C07K14/723 , C07K2319/00 , C07K2319/02 , C12N1/16 , C12N15/81 , C40B30/06 , G01N33/566 , G01N2333/726 , G01N2500/02
摘要: The present invention pertains to novel yeast cells which are useful for the expression of heterologous G protein coupled receptors. The yeast cells of the present invention can be used in screening assays which can be used to screen for modulators of G protein coupled receptors. Specifically, the invention provides novel yeast cells which express a heterologous G protein coupled receptor and mutant and/or chimeric G protein subunit molecules which serve to functionally integrate the heterologous into the pheromone signaling pathway of the yeast cell. The invention also provides for the expression of heterologous G protein coupled receptors which are functionally integrated into the yeast cell membrane using a yeast α factor leader sequence. Drug discovery assays using the subject yeast cells are also provided.
摘要翻译: 本发明涉及可用于表达异源G蛋白偶联受体的新型酵母细胞。 本发明的酵母细胞可用于筛选测定,其可用于筛选G蛋白偶联受体的调节剂。 具体地,本发明提供了表达异源G蛋白偶联受体和突变体和/或嵌合G蛋白亚基分子的新型酵母细胞,其用于将异源整合到酵母细胞的信息素信号通路中。 本发明还提供使用酵母α因子前导序列在功能上整合到酵母细胞膜中的异源G蛋白偶联受体的表达。 还提供使用本发明酵母细胞的药物发现测定。
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公开(公告)号:US07250263B2
公开(公告)日:2007-07-31
申请号:US10752478
申请日:2004-01-05
申请人: Christine A. Klein , Andrew J. M. Murphy , Dana M. Fowlkes , James Broach , John Manfredi , Jeremy Paul , Joshua Trueheart
发明人: Christine A. Klein , Andrew J. M. Murphy , Dana M. Fowlkes , James Broach , John Manfredi , Jeremy Paul , Joshua Trueheart
CPC分类号: C12N15/81 , C07K14/395 , C07K2319/02
摘要: The present invention makes available a rapid, effective assay for screening and identifying pharmaceutically effective compounds that specifically interact with and modulate the activity of a cellular receptor or ion channel. The subject assay enables rapid screening of large numbers of polypeptides in a library to identifying those polypeptides which induce or antagonize receptor bioactivity. The subject assay is particularly amenable for identifying surrogate ligands for orphan receptors.
摘要翻译: 本发明提供用于筛选和鉴定与细胞受体或离子通道的活性特异性相互作用并调节其活性的药学有效化合物的快速有效的测定。 本发明分析使得能够快速筛选文库中的大量多肽以鉴定诱导或拮抗受体生物活性的那些多肽。 主题测定特别适用于鉴定孤儿受体的替代配体。
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公开(公告)号:US06255059B1
公开(公告)日:2001-07-03
申请号:US08582333
申请日:1996-01-17
申请人: Christine A. Klein , Andrew J. M. Murphy , Dana M. Fowlkes , James Broach , John Manfredi , Jeremy Paul , Joshua Trueheart
发明人: Christine A. Klein , Andrew J. M. Murphy , Dana M. Fowlkes , James Broach , John Manfredi , Jeremy Paul , Joshua Trueheart
IPC分类号: C12N510
CPC分类号: C12N15/81 , C07K14/395 , C07K2319/02
摘要: The present invention makes available a rapid, effective assay for screening and identifying pharmaceutically effective compounds that specifically interact with and modulate the activity of a cellular receptor or ion channel. The subject assay enables rapid screening of large numbers of polypeptides in a yeast expression library to identifying those polypeptides which induce or antagonize receptor bioactivity. The subject assay is particularly amenable for identifying surrogate ligands for orphan receptors.
摘要翻译: 本发明提供用于筛选和鉴定与细胞受体或离子通道的活性特异性相互作用并调节其活性的药学有效化合物的快速有效的测定。 本发明分析使得能够在酵母表达文库中快速筛选大量多肽,以鉴定诱导或拮抗受体生物活性的那些多肽。 主题测定特别适用于鉴定孤儿受体的替代配体。
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