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1. 发明申请

US20090257985A1 METHODS OF REGULATING EXPRESSION OF GENES OR OF GENE PRODUCTS USING SUBSTITUTED TETRACYCLINE COMPOUNDS 审中-公开
标题翻译：使用取代的四环素化合物调节基因表达或基因产物的方法
公开(公告)号：US20090257985A1
公开(公告)日：2009-10-15
申请号：US12234265
申请日：2008-09-19
申请人： Mark L. Nelson , Hermann Bujard
发明人： Mark L. Nelson , Hermann Bujard
IPC分类号： A61K35/12 , C12N5/02
CPC分类号： C12N15/635 , A61K31/65 , C12N15/85 , C12N2830/003
摘要： The present invention relates, at least in part, to the use of substituted tetracycline compounds for regulation of expression of nucleic acids operably linked to a tetracycline operator system. The invention pertains to compounds used in a regulatory system which utilizes components of the Tet repressor/operator/inducer system of prokaryotes to regulate gene expression in cells. Use of certain substituted tetracycline compounds, as featured in the methods of the invention, result in improved dose-response results when compared to those for e.g., tetracycline and doxycycline. Certain methods of the invention thus allow for enhanced control of the Tet repressor/operator/inducer system in regulating gene expression in cells.
摘要翻译：本发明至少部分地涉及取代的四环素化合物用于调节与四环素操作系统可操作地连接的核酸表达的用途。本发明涉及利用原核生物的Tet阻遏物/操纵子/诱导物系统的组分调节细胞中基因表达的调节系统中使用的化合物。与本发明方法中所特征的某些取代的四环素化合物的使用相比，与例如四环素和多西环素相比，导致改善的剂量反应结果。因此，本发明的某些方法允许增强Tet阻遏物/操纵子/诱导物系统在调节细胞中的基因表达方面的控制。

2. 发明授权

US06271341B1 Transcriptional activators with graded transactivation potential 有权
标题翻译：具有分级反式激活潜能的转录激活子
公开(公告)号：US06271341B1
公开(公告)日：2001-08-07
申请号：US09577027
申请日：2000-05-23
申请人： Udo Baron , Manfred Gossen , Hermann Bujard
发明人： Udo Baron , Manfred Gossen , Hermann Bujard
IPC分类号： C07K1900
CPC分类号： C07K14/005 , A01K2217/05 , A61K48/00 , C07K2319/00 , C12N15/635 , C12N15/85 , C12N2710/16622 , C12N2830/003 , C12N2830/006
摘要： Transcriptional activators which differ in their activation potential by more than 3 orders of magnitude are provided. The transactivators are fusions between a DNA binding protein (e.g., a Tet repressor) and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). Substitution mutations at amino acid position 442 within the minimal VP16 domain provide transactivators with differing transactivation ability. Moreover, chimeric activation domains comprising both wild type and mutant minimal VP16 domains provide additional variants with differing transactivation ability. Various aspects of the invention pertain to nucleic acid molecules, vectors, host cells, fusion proteins, transgenic and homologous recombinant organisms and methods of regulating gene transcription.
摘要翻译：提供了其激活电位不同于3个数量级的转录激活子。反式激活因子是DNA结合蛋白（例如Tet阻遏物）和衍生自单纯疱疹病毒蛋白16（VP16）的最小转录激活结构域之间的融合物。在最小VP16结构域内的氨基酸位置442处的取代突变体提供具有不同反式激活能力的反式激活因子。此外，包含野生型和突变型最小VP16结构域的嵌合活化域提供了具有不同反式激活能力的另外的变体。本发明的各个方面涉及核酸分子，载体，宿主细胞，融合蛋白，转基因和同源重组生物以及调节基因转录的方法。

3. 发明授权

US5789156A Tetracycline-regulated transcriptional inhibitors 失效
标题翻译：四环素调节的转录抑制剂
公开(公告)号：US5789156A
公开(公告)日：1998-08-04
申请号：US383754
申请日：1995-02-03
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： A01K67/027 , C07K14/035 , C07K14/245 , C12N1/21 , C12N15/63 , C12N15/67 , C12N15/85 , C12Q1/68 , C07H21/04
CPC分类号： C12N15/8509 , C07K14/005 , C07K14/245 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , A01K2217/05 , A01K2217/075 , A01K2217/20 , A01K2267/01 , A01K2267/02 , A01K2267/03 , A01K2267/0393 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N2710/16622 , C12N2830/003 , C12N2830/006 , Y10S435/81
摘要： Nucleic acid molecules and proteins useful for regulating the expression of genes in eukaryotic cells and organisms in a highly controlled manner are disclosed. In the regulatory system of the invention, transcription of a tet operator-linked nucleotide sequence is inhibited by a transcriptional inhibitor fusion protein composed of two polypeptides, a first polypeptide which binds to tet operator sequences either (i) in the absence but not the presence of tetracycline (or an analogue thereof) or (ii) in the presence but not the absence of tetracycline (or an analogue thereof), and a second polypeptide which directly or indirectly inhibits transcription in eukaryotic cells. In one embodiment, the fusion protein comprises a Tet repressor operatively linked to a transcriptional silencer polypeptide. In another embodiment, the fusion protein comprises a mutated Tet repressor operatively linked to a transcriptional silencer polypeptide. The fusion proteins of the invention are useful for reducing the level of transcription of a tet operator-linked target gene. Moreover, the fusion proteins of the invention can be used in combination with tetracycline-regulated transcriptional activator fusion proteins to allow for precise regulation of the expression of one or multiple target genes. Kits including the components of the regulatory system of the invention are also encompassed by the invention.
摘要翻译：公开了用于以高度受控的方式调节真核细胞和生物体中基因表达的核酸分子和蛋白质。在本发明的调节系统中，tet操纵子连接的核苷酸序列的转录受到由两个多肽组成的转录抑制剂融合蛋白的抑制，所述两个多肽是结合tet操纵子序列的第一个多肽，（i）在不存在但不存在的情况下的四环素（或其类似物）或（ii）在存在但不存在四环素（或其类似物）的情况下，以及在真核细胞中直接或间接抑制转录的第二多肽。在一个实施方案中，融合蛋白包含可操作地连接到转录沉默子多肽的Tet阻遏物。在另一个实施方案中，融合蛋白包含与转录沉默子多肽有效连接的突变Tet阻遏物。本发明的融合蛋白可用于降低与tet操纵子连接的靶基因的转录水平。此外，本发明的融合蛋白可以与四环素调节的转录激活物融合蛋白组合使用，以允许精确调节一个或多个靶基因的表达。包括本发明的调节系统的组分的试剂盒也包括在本发明中。

4. 发明授权

US5654168A Tetracycline-inducible transcriptional activator and tetracycline-regulated transcription units 失效
标题翻译：四环素诱导型转录激活因子和四环素调节转录单位
公开(公告)号：US5654168A
公开(公告)日：1997-08-05
申请号：US275876
申请日：1994-07-15
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： A01K67/027 , C07K14/245 , C12N15/63 , C12N15/67 , C12N15/85 , C12P21/00 , C07H21/04 , C12N15/00
CPC分类号： C12N15/8509 , C07K14/245 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , A01K2217/05 , A01K2217/20 , A01K2267/01 , A01K2267/03 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N2830/003 , C12N2830/006
摘要： Nucleic acid molecules and proteins useful for regulating the expression of genes in eukaryotic cells and organisms in an inducible manner are disclosed. In the regulatory system of the invention, transcription of a tet operator-linked nucleotide sequence is stimulated by a transcriptional activator fusion protein composed of two polypeptides, a first polypeptide which binds to tet operator sequences in the presence of tetracycline or a tetracycline analogue and a second polypeptide which directly or indirectly activates transcription in eukaryotic cells. In one embodiment, the fusion protein comprises a mutated Tet repressor operatively linked to a transcriptional activation polypeptide, such as a portion of herpes simplex virus virion protein 16. In the absence of an inducing agent (tetracycline or a tetracycline analogue), transcription of the tet operator-linked nucleotide sequence remains uninduced. In the presence of the inducing agent, transcription of the tet operator-linked nucleotide sequence is stimulated by the transactivator fusion protein of the invention. Novel transcription units which allow for coordinate or independent tetracycline-regulated expression of two or more nucleotide sequences by the transactivator fusion protein of the invention are also disclosed. Kits including the components of the regulatory system of the invention are also encompassed by the invention.
摘要翻译：公开了用于以可诱导的方式调节真核细胞和生物体中基因表达的核酸分子和蛋白质。在本发明的调节系统中，tet操纵子连接的核苷酸序列的转录受到由两个多肽组成的转录激活物融合蛋白的刺激，该多肽是在四环素或四环素类似物存在下与tet操纵子序列结合的第一多肽在真核细胞中直接或间接激活转录的第二多肽。在一个实施方案中，融合蛋白包含可操作地连接到转录激活多肽（例如一部分单纯疱疹病毒颗粒蛋白16）的突变的Tet阻遏物。在不存在诱导剂（四环素或四环素类似物）的情况下， tet操纵子连接的核苷酸序列仍未引起。在诱导剂存在下，通过本发明的反式激活因子融合蛋白刺激tet操纵子连接的核苷酸序列的转录。还公开了通过本发明的反式激活物融合蛋白允许两个或更多个核苷酸序列的协调或独立的四环素调节的表达的新型转录单位。包括本发明的调节系统的组分的试剂盒也包括在本发明中。

5. 发明授权

US09181556B2 Tetracycline inducible transcription control sequence 有权
标题翻译：四环素诱导型转录调控序列
公开(公告)号：US09181556B2
公开(公告)日：2015-11-10
申请号：US13121673
申请日：2009-08-19
申请人： Hermann Bujard , Rainer Loew
发明人： Hermann Bujard , Rainer Loew
IPC分类号： C07H21/04 , C12N15/63 , A61K31/70
CPC分类号： C12N15/635 , C12N2770/40043 , C12N2830/003
摘要： The present invention relates to inducible transcription control sequences for the regulation of gene expression. Specifically, it relates to an transcription control sequence comprising at least two tet operator sequence motifs allowing the binding of tetracycline-dependent transcriptional regulators, wherein each of the tetracycline-dependent transcriptional regulators binds with respect to its neighbor to an opposite face of the DNA helix, and a minimal promoter comprising a TATA box which is linked at its 5′ end to a general transcription factor binding site. Further, the present invention relates to a vector, a host cell, a plant or a non-human transgenic animal comprising the transcription control sequence. Also contemplated is a method for regulating the expression of a nucleic acid sequence being operatively linked to the transcription control sequence of the present invention in a host cell, a plant or a non-human transgenic animal.
摘要翻译：本发明涉及用于调节基因表达的诱导型转录调控序列。具体地说，本发明涉及包含至少两个允许四环素依赖性转录调节因子结合的tet操纵子序列基序的转录控制序列，其中四环素依赖性转录调节因子中的每一个相对于其邻近的DNA螺旋结构，以及包含在其5'末端与一般转录因子结合位点连接的TATA盒的最小启动子。此外，本发明涉及包含转录调控序列的载体，宿主细胞，植物或非人转基因动物。还考虑了在宿主细胞，植物或非人转基因动物中调节与本发明的转录调控序列有效连接的核酸序列的表达的方法。

6. 发明申请

US20100297163A1 RECOMBINANT MALARIA VACCINE 有权
标题翻译：重组疟疾疫苗
公开(公告)号：US20100297163A1
公开(公告)日：2010-11-25
申请号：US12523023
申请日：2008-01-11
申请人： Hermann Bujard , Rolf Lutz , Christian Kauth , Christian Epp , Ute Wöhlbier
发明人： Hermann Bujard , Rolf Lutz , Christian Kauth , Christian Epp , Ute Wöhlbier
IPC分类号： A61K39/015 , A61K38/02 , A61P33/06 , A61P37/04 , C07K14/445 , C12P21/06
CPC分类号： A61K39/015 , C07K14/445 , Y02A50/412
摘要： The present invention refers to a recombinant malaria vaccine and a method for its manufacture.
摘要翻译：本发明涉及重组疟疾疫苗及其制造方法。

7. 发明申请

US20050034183A1 Non-human animal model for analysis of the original and therapy of organ fibrosis 审中-公开
标题翻译：非人动物模型分析原发性和器官性纤维化的治疗
公开(公告)号：US20050034183A1
公开(公告)日：2005-02-10
申请号：US10490885
申请日：2002-09-27
申请人： Rolf Gebhardt , Hermann Bujard
发明人： Rolf Gebhardt , Hermann Bujard
IPC分类号： A01K67/027 , C07K14/495 , C12N15/85 , C12N5/06
CPC分类号： C12N15/8509 , A01K67/0275 , A01K2217/05 , A01K2217/20 , A01K2227/105 , A01K2267/03 , C07K14/495
摘要： The present invention provides a nonhuman animal model for investigating the cause of and the therapy for organ fibrosis. Furthermore, the invention is directed to an animal model comprising a double transgenic, nonhuman animal having the ability to develop a fibrotic phenotype in a conditional and organ-specific manner. Moreover, the invention provides a method for generating the double transgene nonhuman animal.
摘要翻译：本发明提供了用于调查器官纤维化的原因和治疗的非人动物模型。此外，本发明涉及包含具有以条件和器官特异性方式发展纤维化表型的能力的双转基因非人动物的动物模型。此外，本发明提供了产生双转基因非人动物的方法。

8. 发明授权

US06271348B1 Tetracycline-inducible transcriptional inhibitor fusion proteins 有权
标题翻译：四环素诱导型转录抑制剂融合蛋白
公开(公告)号：US06271348B1
公开(公告)日：2001-08-07
申请号：US09489777
申请日：2000-01-24
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： C07K1900
CPC分类号： C07K14/005 , A01K67/0275 , A01K2217/05 , A01K2217/075 , A01K2217/20 , A01K2227/105 , A01K2267/0393 , A61K48/00 , C07K14/245 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N15/62 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , C12N15/8509 , C12N2710/16622 , C12N2830/003 , C12N2830/006 , C12N2830/32
摘要： Methods of regulating gene expression in subjects using tetracycline-responsive fusion proteins are disclosed. In one embodiment, the method involves introducing into a cell the subject a nucleic acid molecule encoding a fusion protein which inhibits transcription, the fusion protein comprising a first polypeptide which binds to a tet operator sequence, operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first polypeptide can binds to a tet operator sequence in the absence, but not the presence, of tetracycline. Alternatively, the first polypeptide can binds to a tet operator sequence in the presence, but not the absence, of tetracycline. In another embodiment, the method of the invention involves obtaining a cell from a subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding an inhibitory fusion protein of the invention to form a modified cell, administering the modified cell to the subject and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecules can be linked or can be separate molecules.
摘要翻译：公开了使用四环素响应融合蛋白调节受试者基因表达的方法。在一个实施方案中，该方法包括向受试者引入编码抑制转录的融合蛋白的核酸分子，该融合蛋白包含与tet操纵子序列结合的第一多肽，该第一多肽与抑制转录的异源第二多肽有效连接在真核细胞中并调节受试者中四环素或其类似物的浓度。在不存在但不存在四环素的情况下，第一种多肽可以结合tet操纵子序列。或者，第一多肽可以在四环素存在下但不存在时与tet操纵子序列结合。在另一个实施方案中，本发明的方法涉及从受试者获得细胞，将在第一核酸分子中引入至少一个tet操纵子序列的第一核酸分子，将编码本发明的抑制性融合蛋白以形成修饰的细胞，向受试者施用修饰的细胞并调节受试者中四环素或其类似物的浓度。第一和第二核酸分子可以连接或可以是分开的分子。

9. 发明授权

US4868111A Gram-positive expression control sequences 失效
标题翻译：革兰氏阳性表达控制序列
公开(公告)号：US4868111A
公开(公告)日：1989-09-19
申请号：US875437
申请日：1986-06-17
申请人： Hermann Bujard , Stuart Le Grice
发明人： Hermann Bujard , Stuart Le Grice
IPC分类号： C12N15/09 , C12N1/20 , C12N1/21 , C12N9/04 , C12N9/06 , C12N9/10 , C12N15/00 , C12N15/70 , C12N15/75 , C12P21/02 , C12R1/125 , C12R1/19 , C12R1/91
CPC分类号： C12N15/70 , C12N15/75 , Y10S435/832 , Y10S435/849
摘要： New gram-positive expression control DNA sequences useful in the expression of pro- or eukaryotic proteins in gram-positive organisms are provided having in the downstream direction of transcription a transcription initiation DNA sequence of gram-negative origin combined with a ribosome binding site-encoding DNA sequence, optionally a foreign gene and a transcription termination sequence. Also described are expression vectors containing these expression control DNA sequences and processes using same for the manufacture of pro- and eukaryotic polypeptides. In addition processes for the manufacture of such gram-positive expression control DNA sequences and such expression vectors are described.
摘要翻译：提供了可用于革兰氏阳性生物体中真核蛋白或真核蛋白表达的新革兰氏阳性表达对照DNA，其在转录的下游方向具有革兰氏阴性来源的转录起始DNA序列与核糖体结合位点编码 DNA序列，任选的外源基因和转录终止序列。还描述了含有这些表达控制DNA序列的表达载体及其用于制备真核和真核多肽的方法。另外还描述了制备这种革兰氏阳性表达控制DNA序列和这些表达载体的方法。

10. 发明授权

US08318185B2 Recombinant malaria vaccine 有权
标题翻译：重组疟疾疫苗
公开(公告)号：US08318185B2
公开(公告)日：2012-11-27
申请号：US12523023
申请日：2008-01-11
申请人： Hermann Bujard , Rolf Lutz , Christian Kauth , Christian Epp , Ute Wöhlbier
发明人： Hermann Bujard , Rolf Lutz , Christian Kauth , Christian Epp , Ute Wöhlbier
IPC分类号： A61K39/015 , C12P21/06
CPC分类号： A61K39/015 , C07K14/445 , Y02A50/412
摘要： The present invention refers to a recombinant malaria vaccine and a method for its manufacture.
摘要翻译：本发明涉及重组疟疾疫苗及其制造方法。

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