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1. 发明授权

US5534418A Controlled expression of recombinant proteins 失效
标题翻译：重组蛋白的控制表达
公开(公告)号：US5534418A
公开(公告)日：1996-07-09
申请号：US166177
申请日：1993-12-10
申请人： Roland M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
发明人： Roland M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
IPC分类号： C12N15/09 , C07K14/00 , C07K14/435 , C07K14/575 , C07K14/635 , C07K14/705 , C07K14/71 , C07K14/72 , C12N5/00 , C12N5/10 , C12N15/00 , C12N15/12 , C12N15/67 , C12P21/02 , C12Q1/68 , C12R1/91 , G01N33/50
CPC分类号： C12N15/67 , C07K14/721 , C07K14/723 , C12Q1/6897
摘要： The present invention provides methods for the controlled production of recombinant proteins in cells. Cells employed in the invention method contain a gene encoding the desired recombinant protein, with transcription of the gene maintained under the control of a transcriptional control element which is activated by a ligand/receptor complex. The ligand/receptor complex is formed when a ligand (which is a hormone or/and analog thereof) is complexed with a receptor (which is a hormone receptor or functional analog thereof which has the transcription activating properties of the receptor). Receptor is produced by the expression of non-endogenous DNA which is also present in the cells used for production of recombinant protein.
摘要翻译：本发明提供了在细胞中控制重组蛋白质生产的方法。本发明方法中使用的细胞含有编码所需重组蛋白的基因，该基因的转录保持在由配体/受体复合物激活的转录控制元件的控制之下。当配体（其是激素或/或其类似物）与受体（其是具有受体的转录激活特性的激素受体或其功能类似物）复合时，形成配体/受体复合物。受体是由用于生产重组蛋白的细胞中也存在的非内源DNA的表达产生的。

2. 发明授权

US06794160B1 Hormone receptor compositions and methods 失效
标题翻译：激素受体组合物和方法
公开(公告)号：US06794160B1
公开(公告)日：2004-09-21
申请号：US08462817
申请日：1995-06-05
申请人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
发明人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
IPC分类号： C12N1512
CPC分类号： C12Q1/6897 , C07K14/721 , C07K14/723 , C12N15/67
摘要： The present invention provides substantially pure DNA's comprised of sequences which encode proteins having the hormone-binding and/or transcription-activating characteristics of a glucocorticoid receptor, a mineralocorticoid receptor, or a thyroid hormone receptor; various plasmids containing receptor sequences which exemplify these DNA's; receptor proteins, including modified functional forms thereof, expressed from these DNA's (or mRNA's); and a bioassay for determining the functionality of a receptor protein. Use of this bioassay has led to the discovery that a necessary and sufficient condition for activation of transcription of a gene (G), whose transcription is activated by hormones complexed with receptors, is the presence of the hormone and its receptor cell (C) where (G) is located. As a result, two new methods for producing desired proteins in genetically engineered cells were discovered.
摘要翻译：本发明提供基本上纯的DNA，其由编码具有糖皮质激素受体，盐皮质激素受体或甲状腺激素受体的激素结合和/或转录激活特征的蛋白质的序列组成; 含有例证这些DNA的受体序列的各种质粒; 受体蛋白，包括其修饰的功能形式，由这些DNA（或mRNA）表达; 以及用于测定受体蛋白质功能的生物测定。这种生物测定的使用已经导致发现，用于激活基因（G）的转录的必需和充分条件（G），其转录由与受体复合的激素激活，是激素及其受体细胞（C）的存在，其中（G）位于。因此，发现了两种在遗传工程细胞中产生所需蛋白质的新方法。

3. 发明授权

US06806359B1 Hormone receptor compositions and methods 失效
标题翻译：激素受体组合物和方法
公开(公告)号：US06806359B1
公开(公告)日：2004-10-19
申请号：US08464574
申请日：1995-06-05
申请人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
发明人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
IPC分类号： C12N1512
CPC分类号： C12Q1/6897 , C07K14/721 , C07K14/723 , C12N15/67
摘要： The present invention provides substantially pure DNA's comprised of sequences which encode proteins having the hormone-binding and/or transcription-activating characteristics of a glucocorticoid receptor, a mineralocorticoid receptor, or a thyroid hormone receptor. The invention also provides various plasmids containing receptor sequences which exemplify the DNA's of the invention. The invention further provides receptor proteins, including modified functional forms thereof, expressed from the DNA's (or mRNA's) of the invention. In addition to novel receptor DNA, RNA and protein compositions, the present invention involves a bioassay for determining the functionality of a receptor protein. By using our bioassay system we have discovered that a necessary and sufficient condition for activation of transcription of a gene (G), whose transcription is activated by hormones complexed with receptors, is the presence of the hormone and its receptor in the cell (C) where (G) is located. As a result of that discovery we have also invented new methods for producing desired proteins in genetically engineered cells. Two of these methods are methods of the present invention. The first is a method for inducing transcription of a gene whose transcription is activated by hormones complexed with the receptors. The second is a method for engineering a cell and increasing and controlling production of a protein encoded by a gene whose transcription is activated by hormones complexed with receptor proteins.
摘要翻译：本发明提供基本上纯的DNA，其由编码具有糖皮质激素受体，盐皮质激素受体或甲状腺激素受体的激素结合和/或转录激活特征的蛋白质的序列组成; 含有例证这些DNA的受体序列的各种质粒; 受体蛋白，包括其修饰的功能形式，由DNA（或mRNA）表达; 以及用于测定受体蛋白质功能的生物测定。这种生物测定的使用已经导致发现，用于激活基因（G）的转录的必需和充分条件（G），其转录由与受体复合的激素激活，是激素及其受体细胞（C）的存在，其中（G）位于。因此，发现了两种在遗传工程细胞中产生所需蛋白质的新方法。

4. 发明授权

US5597705A DNA encoding thyroid hormone receptor compositions and methods 失效
标题翻译：编码甲状腺激素受体组成和方法的DNA
公开(公告)号：US5597705A
公开(公告)日：1997-01-28
申请号：US165708
申请日：1993-12-10
申请人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
发明人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
IPC分类号： C12N15/09 , C07K14/00 , C07K14/435 , C07K14/575 , C07K14/635 , C07K14/705 , C07K14/71 , C07K14/72 , C12N5/00 , C12N5/10 , C12N15/00 , C12N15/12 , C12N15/67 , C12P21/02 , C12Q1/68 , C12R1/91 , G01N33/50 , C22N15/62
CPC分类号： C12N15/67 , C07K14/721 , C07K14/723 , C12Q1/6897
摘要： The present invention provides a recombinant expression system for production of functional thyroid hormone receptor protein(s). The invention also provides a method to produce thyroid hormone receptor protein(s) by culturing the cells of the invention recombinant expression system. Also provided are thyroid hormone receptor protein(s) produced by the invention method. In addition, the present invention provides recombinant DNAs comprised of sequences which encode proteins having the hormone-binding and/or transcription-activating characteristics of a thyroid hormone receptor. The invention also provides various plasmids containing receptor sequences which exemplify the DNAs of the invention. The invention further provides complementary mRNAs, cells transformed with invention DNAs, and nucleic acid probes derived from invention DNAs.
摘要翻译：本发明提供了用于产生功能性甲状腺激素受体蛋白质的重组表达系统。本发明还提供了通过培养本发明的重组表达系统的细胞产生甲状腺激素受体蛋白的方法。还提供了通过本发明方法产生的甲状腺激素受体蛋白。此外，本发明提供由编码具有甲状腺激素受体的激素结合和/或转录激活特征的蛋白质的序列组成的重组DNA。本发明还提供了含有受体序列的各种质粒，其例证了本发明的DNA。本发明还提供了补体mRNA，用本发明DNA转化的细胞，以及衍生自本发明DNA的核酸探针。

5. 发明授权

US5312732A Hormone receptor compositions and methods 失效
标题翻译：激素受体组合物和方法
公开(公告)号：US5312732A
公开(公告)日：1994-05-17
申请号：US667602
申请日：1991-03-07
申请人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
发明人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
IPC分类号： C12N15/09 , C07K14/00 , C07K14/435 , C07K14/575 , C07K14/635 , C07K14/705 , C07K14/71 , C07K14/72 , C12N5/00 , C12N5/10 , C12N15/00 , C12N15/12 , C12N15/67 , C12P21/02 , C12Q1/68 , C12R1/91 , G01N33/50 , C12N15/63
CPC分类号： C12N15/67 , C07K14/721 , C07K14/723 , C12Q1/6897
摘要： The present invention provides substantially pure DNA's comprised of sequences which encode proteins having the hormone-binding and/or transcription-activating characteristics of a glucocorticoid receptor, a mineralocorticoid receptor, or a thyroid hormone receptor. The invention also provides various plasmids containing receptor sequences which exemplify the DNA's of the invention. The invention further provides receptor proteins, including modified functional forms thereof, expressed from the DNA's (or mRNA's) of the invention. In addition to the novel receptor DNA, RNA and protein compositions, the present invention involves a bioassay for determining the functionality of a receptor protein. By using our bioassay system we have discovered that a necessary and sufficient condition for activation of transcription of a gene (G), whose transcription is activated by hormones complexed with receptors, is the presence of the hormone and its receptor in the cell (C) where (G) is located. As a result of that discovery we have also invented new methods for producing desired proteins in genetically engineered cells. Two of these methods are methods of the present invention. The first is a method for inducing transcription of a gene whose transcription is activated by hormones complexed with the receptors. The second is a method for engineering a cell and increasing and controlling production of a protein encoded by a gene whose transcription is activated by hormones complexed with receptor proteins.
摘要翻译：本发明提供了基本上纯的DNA，其由编码具有糖皮质激素受体，盐皮质激素受体或甲状腺激素受体的激素结合和/或转录激活特征的蛋白质的序列组成。本发明还提供了含有受体序列的各种质粒，其例证了本发明的DNA。本发明还提供了从本发明的DNA（或mRNA）表达的受体蛋白，包括其修饰的功能形式。除了新的受体DNA，RNA和蛋白质组合物之外，本发明涉及用于测定受体蛋白质的功能的生物测定。通过使用我们的生物测定系统，我们已经发现，激活基因（G）的转录的必要和充分条件（G），其转录由与受体复合的激素激活，是细胞中激素及其受体的存在（C）其中（G）位于。作为这一发现的结果，我们还发明了用于在遗传工程细胞中产生所需蛋白质的新方法。这些方法中的两个是本发明的方法。第一种是诱导转录由与受体复合的激素激活的基因的转录的方法。第二种是用于工程化细胞并增加和控制由受体与受体蛋白质复合的激素激活转录的基因编码的蛋白质的产生的方法。

6. 发明授权

US5071773A Hormone receptor-related bioassays 失效
标题翻译：激素受体相关生物多样性
公开(公告)号：US5071773A
公开(公告)日：1991-12-10
申请号：US108471
申请日：1987-10-20
申请人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
发明人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Vincent Giguere , Jeffrey Arriza , Catherine C. Thompson , Estelita S. Ong
IPC分类号： C12N15/09 , C07K14/00 , C07K14/435 , C07K14/575 , C07K14/635 , C07K14/705 , C07K14/71 , C07K14/72 , C12N5/00 , C12N5/10 , C12N15/00 , C12N15/12 , C12N15/67 , C12P21/02 , C12Q1/68 , C12R1/91 , G01N33/50
CPC分类号： C12N15/67 , C07K14/721 , C07K14/723 , C12Q1/6897
摘要： The present invention discloses two hormone receptor-related bioassays. The first bioassay is useful for determining whether a protein suspected of being a hormone receptor has transcription-activating properties of a hormone receptor. The second bioassay is useful for evaluating whether compounds are functional ligands for receptor proteins. According to the first bioassay, cells that contain non-endogenous DNA which expresses a protein suspected of being a hormone receptor and which contain a DNA sequence encoding an operative hormone responsive promoter/enhancer element linked to an operative reporter gene, are cultured, the culturing being conducted in a culture medium containing a known hormone, or an analog thereof. The cultured cells are then monitored for induction of the product of the reporter gene as an indication of functional transcription-activating binding between the hormone or hormone analog and the protein suspected of being a hormone receptor. According to the second bioassay, cells that contain non-endogenous DNA which expresses hormone receptor or a functional engineered or modified form thereof, and which also contain a DNA sequence encoding an operative hormone responsive promoter/enhancer element linked to an operative reporter gene, are cultured, the culturing being conducted in culture medium containing at least one compound whose ability to functionally bind the receptor protein is sought to be determined. The cultured cells are then monitored for induction of the product of the report gene as an indicator of functional binding between the compound and the receptor.
摘要翻译：本发明公开了两种与激素受体相关的生物测定。第一种生物测定可用于确定怀疑是激素受体的蛋白质是否具有激素受体的转录激活特性。第二个生物测定可用于评估化合物是否是受体蛋白质的功能性配体。根据第一生物测定法，培养含有表达怀疑是激素受体的蛋白质并含有编码与操作性报告基因连接的操作性激素反应性启动子/增强子元件的DNA序列的非内源性DNA的细胞，培养在含有已知激素或其类似物的培养基中进行。然后监测培养的细胞以诱导报告基因的产物，作为激素或激素类似物与怀疑是激素受体的蛋白质之间的功能性转录激活结合的指示。根据第二生物测定，含有表达激素受体的非内源性DNA或其功能性工程化或修饰形式的细胞，并且还包含编码与有效的报告基因连接的操作性激素反应性启动子/增强子元件的DNA序列的细胞是培养时，寻求在含有至少一种能够与受体蛋白质功能结合的化合物的培养基中进行培养。然后监测培养的细胞以诱导报告基因的产物，作为化合物和受体之间功能性结合的指标。

7. 发明授权

US07214511B1 Hormone receptor compositions and methods 失效
标题翻译：激素受体组合物和方法
公开(公告)号：US07214511B1
公开(公告)日：2007-05-08
申请号：US08464262
申请日：1995-06-05
申请人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Estelita S. Ong
发明人： Ronald M. Evans , Cary A. Weinberger , Stanley M. Hollenberg , Estelita S. Ong
IPC分类号： C12N15/12
CPC分类号： C12Q1/6897 , C07K14/721 , C07K14/723 , C12N15/67
摘要： In accordance with the present invention, there are provided expression systems for the production of function glucocorticoid receptor proteins, methods for the recombinant production of glucocorticoid receptor proteins as well as sequences encoding novel members of the steroid/thyroid hormone superfamily of receptors (e.g., glucocorticoid receptor). Invention expression systems comprise host cells containing DNA encoding functional glucocorticoid receptor proteins, wherein the DNA is operably linked to control sequences compatible with host cells, thereby enabling the expression of functional receptor proteins. The invention method is carried out by culturing such recombinant host cells, and recovering the functional glucocorticoid receptor proteins produced thereby. In accordance with the present invention, there are also provided substantially pure DNA's comprised of sequences which encode members of the steroid/thyroid hormone receptor superfamily having the hormone-binding and/or transcription-activating characteristics of a glucocorticoid receptor. The invention also provides various plasmids containing receptor sequences which exemplify the DNA's of the invention. The invention further provides receptor proteins, including modified functional forms thereof, expressed from the DNA's (or mRNA's) of the invention.
摘要翻译：根据本发明，提供了用于产生功能性糖皮质激素受体蛋白质的表达系统，重组生产糖皮质激素受体蛋白质的方法以及编码受体类固醇/甲状腺激素超家族的新成员的序列（例如，糖皮质激素受体）。本发明表达系统包含含有编码功能性糖皮质激素受体蛋白质的DNA的宿主细胞，其中所述DNA可操作地连接到与宿主细胞相容的控制序列，从而使得能够表达功能性受体蛋白质。本发明方法是通过培养这样的重组宿主细胞进行的，并且回收由此产生的功能性糖皮质激素受体蛋白质。根据本发明，还提供了基本上纯的DNA，其由编码具有糖皮质激素受体的激素结合和/或转录激活特性的类固醇/甲状腺激素受体超家族成员的序列组成。本发明还提供了含有受体序列的各种质粒，其例证了本发明的DNA。本发明还提供了从本发明的DNA（或mRNA）表达的受体蛋白，包括其修饰的功能形式。

8. 发明授权

US5298429A Bioassay for identifying ligands for steroid hormone receptors 失效
标题翻译：用于鉴定类固醇激素受体配体的生物测定
公开(公告)号：US5298429A
公开(公告)日：1994-03-29
申请号：US807135
申请日：1991-12-10
申请人： Ronald M. Evans , Stanley M. Hollenberg , Vincent Giguere
发明人： Ronald M. Evans , Stanley M. Hollenberg , Vincent Giguere
IPC分类号： C12N15/09 , C07K14/00 , C07K14/435 , C07K14/575 , C07K14/635 , C07K14/705 , C07K14/71 , C07K14/72 , C12N5/00 , C12N5/10 , C12N15/00 , C12N15/12 , C12N15/67 , C12P21/02 , C12Q1/68 , C12R1/91 , G01N33/50 , G01N30/00
CPC分类号： C12N15/67 , C07K14/721 , C07K14/723 , C12Q1/6897
摘要： Bioassays are disclosed which are useful for determining whether a compound is a hormone receptor agonist (i.e., is capable of promoting the transcription-activation activities of such receptors) or a hormone receptor antagonist (i.e., is capable of blocking the transcription-activation activities of such receptors). The invention bioassay is conducted by culturing test cells in the presence of at least one compound whose ability to function as a ligand for said receptor protein (or functional engineered or modified forms thereof) is sought to be determined. Alternatively, test cells are cultured in medium containing increasing concentrations of at least one compound whose ability to inhibit the transcription activation activity of hormone receptor agonists is sought to be determined, and a fixed concentration of at least one agonist for the receptor protein. Test cells employed in the practice of the present invention contain non-endogenous DNA which expresses hormone receptor (or functional modified forms thereof) and a DNA sequence encoding a hormone response element operatively linked to a reporter gene. The cultured cells are monitored for evidence of transcription of the reporter gene as a function of the concentration of test compound in the culture medium. The variation in transcription levels of the reporter gene as a function of concentration of test compound indicates the ability of test compound to promote or inhibit activation of transcription.
摘要翻译：公开了可用于确定化合物是激素受体激动剂（即能够促进此类受体的转录激活活性）还是激素受体拮抗剂的生物测定（即，能够阻断化合物的转录活化活性这样的受体）。本发明生物测定通过在至少一种化合物的存在下培养测试细胞来进行，该化合物的功能是用作所述受体蛋白质的配体（或其功能性工程化或修饰形式）。或者，将测试细胞在含有增加浓度的至少一种其抑制激素受体激动剂的转录激活活性的能力的化合物的培养基中培养，并测定受体蛋白质的至少一种激动剂的固定浓度。在本发明的实践中使用的测试细胞含有表达激素受体（或其功能性修饰形式）的非内源性DNA和编码与报告基因有效连接的激素应答元件的DNA序列。监测培养的细胞作为培养基中测试化合物浓度的函数的报告基因转录的证据。作为测试化合物浓度的函数的报告基因的转录水平的变化表明测试化合物促进或抑制转录激活的能力。

9. 发明授权

US5217867A Receptors: their identification, characterization, preparation and use 失效
标题翻译：受体：他们的鉴定，表征，制备和使用
公开(公告)号：US5217867A
公开(公告)日：1993-06-08
申请号：US278614
申请日：1988-11-30
申请人： Ronald M. Evans , Stanley M. Hollenberg
发明人： Ronald M. Evans , Stanley M. Hollenberg
IPC分类号： C12N15/09 , C07H21/04 , C07K14/435 , C07K14/705 , C07K14/72 , C12N5/10 , C12N15/12 , C12N15/85 , C12P21/00 , C12Q1/48 , C12Q1/68 , C12R1/91 , G01N33/566 , G01N33/574 , G01N33/74
CPC分类号： G01N33/574 , C07K14/721 , C12N15/85 , C12Q1/6897 , G01N33/566 , G01N33/743 , C07K2319/00 , C07K2319/61 , C07K2319/715 , C07K2319/81 , C12N2830/002 , C12N2830/15 , C12N2830/60 , G01N2333/723 , G01N2500/00
摘要： Disclosed are novel hormone and hormone-like receptors wherein the trans-activation transcription domain(s) is modified in terms of position and/or copy number or otherwise versus parent receptor, such novel receptors having increased trans-activation transcription activation properties surprisingly superior to the parent receptor. Also disclosed are the recombinant methods and means for preparing such receptors and assays based upon the use of such receptors for screening and identifying putative materials that can affect such receptors and/or for the expression via induced transcription of a reporter or other desired, preferably heterologous gene or DNA product.

10. 发明授权

US5310662A Receptors: their identification, characterization, preparation and use 失效
标题翻译：受体：其鉴别，鉴定，准备和使用
公开(公告)号：US5310662A
公开(公告)日：1994-05-10
申请号：US73928
申请日：1993-06-08
申请人： Ronald M. Evans , Stanley M. Hollenberg
发明人： Ronald M. Evans , Stanley M. Hollenberg
IPC分类号： C12N15/09 , C07H21/04 , C07K14/435 , C07K14/705 , C07K14/72 , C12N5/10 , C12N15/12 , C12N15/85 , C12P21/00 , C12Q1/48 , C12Q1/68 , C12R1/91 , G01N33/566 , G01N33/574 , G01N33/74 , C12N15/62 , C12N15/63
CPC分类号： G01N33/574 , C07K14/721 , C12N15/85 , C12Q1/6897 , G01N33/566 , G01N33/743 , C07K2319/00 , C07K2319/61 , C07K2319/715 , C07K2319/81 , C12N2830/002 , C12N2830/15 , C12N2830/60 , G01N2333/723 , G01N2500/00
摘要： Disclosed are novel hormone and hormone-like receptors Wherein the trans-activation transcription domain(s) is modified in terms of position and/or copy number or otherwise versus parent receptor, such novel receptors having increased trans-activation transcription activation properties surprisingly superior to the parent receptor. Also disclosed are the recombinant methods and means for preparing such receptors and assays based upon the use of such receptors for screening and identifying putative materials that can affect such receptors and/or for the expression via induced transcription of a reporter or other desired, preferably heterologous gene or DNA product.
摘要翻译：公开了新颖的激素和激素样受体，其中反式激活转录结构域在位置和/或拷贝数或其他方面根据亲本受体进行修饰，这种具有增加的反式激活转录激活特性的新受体惊人地优于亲本受体。还公开了用于制备这种受体的重组方法和方法以及基于这种受体用于筛选和鉴定可影响此类受体的推定材料和/或通过报告物的诱导转录或其他所需的优选异源基因或DNA产物。

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