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1. 发明申请

US20080268475A1 YEAST CELLS EXPRESSING MODIFIED G PROTEINS AND METHODS OF USE THEREFOR 审中-公开
标题翻译： YEAST CELLS EXPRESSING MODIFIED G蛋白及其使用方法
公开(公告)号：US20080268475A1
公开(公告)日：2008-10-30
申请号：US12106678
申请日：2008-04-21
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： G01N33/569 , C12N1/16 , C40B30/04
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。

2. 发明授权

US07105309B2 Yeast cells engineered to produce pheromone system protein surrogates and uses therefor 失效
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.

3. 发明授权

US07361498B2 Yeast cells expressing modified G proteins and methods of use therefor 失效
标题翻译：酵母细胞表达修饰的G蛋白及其使用方法
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
IPC分类号： C12N1/20 , C07H17/00 , C07K14/00
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。

4. 发明授权

US06864060B1 Yeast cells expressing modified G proteins and methods of use therefor 失效
标题翻译：酵母细胞表达修饰的G蛋白及其使用方法
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。

5. 发明授权

US6001553A Functional expression of mammalian adenylyl cyclase in yeast 失效
标题翻译：酵母中哺乳动物腺苷酸环化酶的功能表达
公开(公告)号：US6001553A
公开(公告)日：1999-12-14
申请号：US732218
申请日：1997-01-14
申请人： James R. Broach , John P. Manfredi , Joshua Trueheart
发明人： James R. Broach , John P. Manfredi , Joshua Trueheart
IPC分类号： G01N33/573 , C12N1/19 , C12N9/88 , C12N15/09 , C12N15/60 , C12Q1/02 , C12Q1/00 , C12N1/14
CPC分类号： C12N9/88
摘要： Mammalian adenylyl cyclases are functionally expressed in yeast cells. The yeast cells may be used to screen for inhibitors or activators of the adenylyl cyclase, or of a regulator of adenylyl cyclase which is functionally co-expressed in the yeast cell. Methods of identifying such inhibitors, activators and regulators are also disclosed.
摘要翻译： PCT No.PCT / US95 / 05149 Sec。 371日期1997年1月14日 102（e）日期1997年1月14日PCT 1995年4月26日PCT PCT。第WO95 / 30012号公报日期：1995年11月9日玛氏腺苷酸环化酶在酵母细胞中功能表达。酵母细胞可用于筛选腺苷酸环化酶的抑制剂或活化剂，或在酵母细胞中功能共表达的腺苷酸环化酶的调节剂。还公开了鉴定这种抑制剂，活化剂和调节剂的方法。

6. 发明授权

US07273747B2 Cell having amplified signal transduction pathway responses and uses therefor 失效
标题翻译：细胞具有扩增的信号转导途径反应和用途
公开(公告)号：US07273747B2
公开(公告)日：2007-09-25
申请号：US10267074
申请日：2002-10-08
申请人： John P. Manfredi , Benjamin K. Benton , Meng-Yu Wu
发明人： John P. Manfredi , Benjamin K. Benton , Meng-Yu Wu
IPC分类号： C12N1/19 , C12N15/81 , G01N33/554
CPC分类号： C12N15/81 , C07K14/723
摘要： The invention provides recombinant cells that have been engineered such that ligand stimulation of a receptor expressed by the cells leads to amplified signal transduction responses. In one embodiment, the receptor-expressing cells have been engineered to carry a heterologous DNA construct comprising a gene encoding a protein that activates the signal transduction pathway, which gene is operatively linked to a promoter that is responsive to activation of the signal transduction pathway. Stimulation of the receptor by a ligand leads to expression of the heterologous DNA construct encoding the protein that activates the signal transduction pathway such that signals generated by ligand binding to the receptor are amplified. Preferred cells are yeast cells expressing heterologous G protein coupled receptors functionally coupled to the yeast pheromone response pathway and overexpressing Ste5p, Ste4p, Ste12p, Ste11p or a dominant truncation allele of Ste20 via a pheromone-responsive promoter. The invention further provides cells expressing a heterologous receptor, wherein an endogenous gene encoding a protein that negatively regulates an endogenous signal transduction pathway is mutated to render the protein nonfunctional such that signals generated by ligand binding to the receptor are amplified. The invention further provides cells expressing a heterologous receptor, wherein an endogenous gene encoding a protein that positively regulates an endogenous signal transduction pathway is mutated to a supersensitive form such that the sensitivity of the pathway to ligand stimulation is increases. Methods of using the cells of the invention to identify receptor modulators are also provided.
摘要翻译：本发明提供重组细胞，这些细胞已经被改造成使得由细胞表达的受体的配体刺激导致扩增的信号转导反应。在一个实施方案中，受体表达细胞已被改造以携带包含编码活化信号转导途径的蛋白质的基因的异源DNA构建体，该基因可操作地连接于对信号转导途径的激活有反应的启动子。通过配体刺激受体导致编码激活信号转导途径的蛋白质的异源DNA构建体的表达，使得通过配体结合受体产生的信号被扩增。优选的细胞是表达与酵母信息素反应途径功能耦合的异源G蛋白偶联受体的酵母细胞，并且通过信息素反应启动子过表达Ste5p，Ste4p，Ste12p，Ste11p或Ste20的显性截短等位基因。本发明还提供了表达异源受体的细胞，其中编码负调节内源信号转导途径的蛋白质的内源基因被突变以使蛋白质无功能化，使得通过配体结合受体产生的信号被扩增。本发明进一步提供表达异源受体的细胞，其中编码将内源性信号转导途径正调节的蛋白质的内源基因突变为超敏感形式，使得该途径对配体刺激的敏感性增加。还提供了使用本发明的细胞鉴定受体调节剂的方法。

7. 发明申请

US20120325152A1 METHODS AND COMPOSITIONS FOR NEMATODE GROWTH 审中-公开
标题翻译： NEMATODE GROWTH的方法和组合物
公开(公告)号：US20120325152A1
公开(公告)日：2012-12-27
申请号：US13531039
申请日：2012-06-22
申请人： Christopher E. Hopkins , John P. Manfredi
发明人： Christopher E. Hopkins , John P. Manfredi
IPC分类号： A01K67/033 , A23L1/48
CPC分类号： A01K67/033
摘要： Compositions for nematode growth and methods for using the same for nematode viability are described. The compositions contain a solid matrix material that is dispersed in a liquid medium. The solid matrix can be immersed, bathed, and/or interspersed in a liquid medium to form a composite culture that is capable of maintaining or growing nematodes at unexpectedly high densities and/or with nominal stress pathway activity. Other embodiments are described.
摘要翻译：描述了用于线虫生长的组合物及其用于线虫活力的方法。组合物含有分散在液体介质中的固体基质材料。可以将固体基质浸入，洗涤和/或散布在液体培养基中以形成能够以意想不到的高密度和/或具有标称应力通路活性保持或生长线虫的复合培养物。描述其他实施例。

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