公开(公告)号：US5814618A

公开(公告)日：1998-09-29

申请号：US485978

申请日：1995-06-07

申请人： Hermann Bujard ,  Manfred Gossen

发明人： Hermann Bujard ,  Manfred Gossen

IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C07K14/035 ,  C07K14/195 ,  C07K14/245 ,  C07K19/00 ,  C12N15/11 ,  C12N15/63 ,  C12N15/67 ,  C12N15/85 ,  C12N15/00

CPC分类号： C07K14/005 ,  A01K67/0275 ,  C07K14/245 ,  C12N15/62 ,  C12N15/63 ,  C12N15/635 ,  C12N15/67 ,  C12N15/85 ,  C12N15/8509 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2217/20 ,  A01K2227/105 ,  A01K2267/0393 ,  A61K48/00 ,  C07K2319/00 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  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。

公开(公告)号：US5650298A

公开(公告)日：1997-07-22

申请号：US260452

申请日：1994-06-14

申请人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

发明人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C12N5/08 ,  C12N5/10 ,  C12N15/00 ,  C12N15/09 ,  C12N15/31 ,  C12N15/62 ,  C12N15/63 ,  C12N15/85 ,  C12R1/91 ,  C12P21/00 ,  C07H21/04

CPC分类号： A01K67/0275 ,  C12N15/62 ,  C12N15/635 ,  C12N15/85 ,  A01K2217/05 ,  A01K2217/075 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2830/006

摘要： Transgenic animals carrying two transgenes, the first coding for a transactivator fusion protein comprising a tet repressor and a polypeptide which directly or indirectly activates in eucaryotic cells, and the second comprising a gene operably linked to a minimal promoter operably linked to at least one tet operator sequence, are disclosed. Isolated DNA molecules (e.g., targeting vectors) for integrating a polynucleotide sequence encoding a transactivator of the invention at a predetermined location within a second target DNA molecule by homologous recombination are also disclosed. Transgenic animals having the DNA molecules of the invention integrated at a predetermined location in a chromosome by homologous recombination are also encompassed by the invention. Methods to regulate the expression of a tet operator linked-gene of interest by administering tetracycline or a tetracycline analogue to an animal of the invention are also disclosed. The regulatory system of the invention allows for conditional inactivation or modulation of expression of a gene of interest in a host cell or animal.

摘要翻译： 携带两个转基因的转基因动物，第一编码包含直接或间接在真核细胞中活化的tet阻遏物和多肽的反式激活因子融合蛋白，第二编码包含与可操作地连接至少一个tet操纵子的最小启动子可操作地连接的基因 序列。 还公开了通过同源重组将编码本发明的反式激活物的多核苷酸序列整合到第二靶DNA分子内的预定位置的分离的DNA分子（例如靶向载体）。 通过同源重组将具有整合在染色体中预定位置的本发明的DNA分子的转基因动物也包括在本发明中。 还公开了通过向本发明的动物施用四环素或四环素类似物来调节所述目的基因的表达的方法。 本发明的调节系统允许在宿主细胞或动物中有条件地失活或调控感兴趣的基因的表达。

公开(公告)号：US5589362A

公开(公告)日：1996-12-31

申请号：US485971

申请日：1995-06-07

申请人： Hermann Bujard ,  Manfred Gossen ,  Wolfgang Hillen ,  Vera Helbl ,  Dirk Schnappinger

发明人： Hermann Bujard ,  Manfred Gossen ,  Wolfgang Hillen ,  Vera Helbl ,  Dirk Schnappinger

IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C07K14/035 ,  C07K14/195 ,  C07K14/245 ,  C07K19/00 ,  C12N15/11 ,  C12N15/63 ,  C12N15/67 ,  C12N15/85 ,  C12P21/00 ,  C07H21/04 ,  C12N15/31

CPC分类号： A01K67/0275 ,  C07K14/005 ,  C07K14/245 ,  C12N15/63 ,  C12N15/635 ,  C12N15/67 ,  C12N15/85 ,  C12N15/8509 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2217/20 ,  A01K2227/105 ,  A01K2267/0393 ,  A61K48/00 ,  C07K2319/00 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2710/16622 ,  C12N2830/003 ,  C12N2830/006 ,  C12N2830/30 ,  C12N2830/32

摘要： Isolated nucleic acid molecules encoding fusion proteins which regulate transcription in eukaryotic cells are disclosed. The fusion proteins of the invention comprise a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position +4 or +6, operatively linked to a polypeptide which regulates transcription in eukaryotic cells. Methods for regulating transcription of a tet operator-linked gene in a cell are also provided. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a fusion protein which regulates transcription, the fusion protein comprising a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position +4 or +6, operatively linked to a polypeptide which regulates transcription in eukaryotic cells, and modulating the concentration of a tetracycline, or analogue thereof, in contact with the cell.

摘要翻译： 公开了编码在真核细胞中调节转录的融合蛋白的分离的核酸分子。 本发明的融合蛋白包含Tet阻遏物，其具有赋予融合蛋白的至少一个氨基酸突变，其结合具有与多肽有效连接的位置+4或+6的核苷酸取代的B类tet操纵子序列的能力 其调控真核细胞中的转录。 还提供了调节细胞中tet操纵子连接基因转录的方法。 在一个实施方案中，该方法包括将编码调节转录的融合蛋白的核酸分子导入细胞，所述融合蛋白包含具有至少一个氨基酸突变的Tet阻遏物，赋予融合蛋白具有结合B类的能力 tet操纵子序列在+4或+6位具有核苷酸取代，可操作地连接到调节真核细胞转录的多肽，并调节与细胞接触的四环素或其类似物的浓度。

公开(公告)号：US06914124B2

公开(公告)日：2005-07-05

申请号：US09777317

申请日：2001-02-05

申请人： Hermann Bujard ,  Manfred Gossen

发明人： Hermann Bujard ,  Manfred Gossen

IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C12N5/08 ,  C12N5/10 ,  C12N15/00 ,  C12N15/09 ,  C12N15/31 ,  C12N15/62 ,  C12N15/63 ,  C12N15/85 ,  C12R1/91

CPC分类号： A01K67/0275 ,  A01K2217/05 ,  A01K2217/075 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N15/62 ,  C12N15/635 ,  C12N15/85 ,  C12N2830/006

摘要： A method for regulating expression of a tet operator-linked gene in a cell of a subject is disclosed. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a tetracycline-controllable transactivator (tTA), the tTA comprising a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. Alternatively, in another embodiment, the method involves obtaining the cell from the 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 a tTA, 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 molecule can be within a single molecule (e.g., in the same vector) or on separate molecules.

摘要翻译： 公开了一种用于调节受试者细胞中tet操纵子连接基因表达的方法。 在一个实施方案中，所述方法包括将编码四环素可控反式激活因子（tTA）的核酸分子引入细胞，所述tTA包含可操作地连接到直接或间接激活真核细胞中的转录的多肽的Tet阻遏物; 并调节受试者中四环素或其类似物的浓度。 或者，在另一个实施方案中，所述方法包括从受试者获得细胞，向细胞中引入第一核酸分子，该第一核酸分子可操作地将基因连接至至少一个tet操纵子序列，将编码tTA的第二核酸分子 ，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在单分子内（例如，在同一载体中）或分开的分子上。

公开(公告)号：US06242667B1

公开(公告)日：2001-06-05

申请号：US09161902

申请日：1998-09-28

申请人： Hermann Bujard ,  Manfred Gossen

发明人： Hermann Bujard ,  Manfred Gossen

IPC分类号： C12N1509

CPC分类号： C07K14/005 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2217/20 ,  A01K2227/105 ,  A01K2267/01 ,  A01K2267/02 ,  A01K2267/03 ,  A01K2267/0393 ,  A61K48/00 ,  C07K14/245 ,  C07K2319/00 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N15/85 ,  C12N15/8509 ,  C12N2710/16622 ,  C12N2800/30 ,  C12N2830/00 ,  C12N2830/003 ,  C12N2830/006 ,  C12N2830/32

摘要： Transgenic animals carrying a transgene comprising a nucleic acid molecule encoding protein 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 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 operatively linked to a second polypeptide activates transcription in eukaryotic cells. In a preferred embodiment, the transgene encoding the transcriptional activator fusion protein is integrated at a predetermined location within the chromosome of the transgenic animal.

摘要翻译： 公开了携带转基因的转基因，其包含编码可用于以高度受控的方式调控真核细胞和生物体中的基因表达的蛋白质的核酸分子。 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，所述两个多肽是在与第二个多肽有效连接的四环素存在下结合tet操纵子序列的第一多肽 在真核细胞中激活转录。 在优选的实施方案中，编码转录激活物融合蛋白的转基因整合在转基因动物的染色体内的预定位置。

公开(公告)号：US6087166A

公开(公告)日：2000-07-11

申请号：US888080

申请日：1997-07-03

申请人： Udo Baron ,  Manfred Gossen ,  Hermann Bujard

发明人： Udo Baron ,  Manfred Gossen ,  Hermann Bujard

IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/035 ,  C07K14/72 ,  C07K19/00 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N15/09 ,  C12N15/63 ,  C12N15/85 ,  C12N1/00 ,  C12N15/62

CPC分类号： C07K14/005 ,  C12N15/635 ,  C12N15/85 ,  A01K2217/05 ,  A61K48/00 ,  C07K2319/00 ,  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。

公开(公告)号：US5922927A

公开(公告)日：1999-07-13

申请号：US897719

申请日：1997-07-21

申请人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

发明人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C12N5/08 ,  C12N5/10 ,  C12N15/00 ,  C12N15/09 ,  C12N15/31 ,  C12N15/62 ,  C12N15/63 ,  C12N15/85 ,  C12R1/91 ,  C12N5/00

CPC分类号： A01K67/0275 ,  C12N15/62 ,  C12N15/635 ,  C12N15/85 ,  A01K2217/05 ,  A01K2217/075 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2830/006

摘要： Transgenic mice carrying two transgenes, the first coding for a transactivator fusion protein comprising a tet repressor and a polypeptide which directly or indirectly activates in eucaryotic cells, and the second comprising a gene operably linked to a minimal promoter operably linked to at least one tet operator sequence, are disclosed. Isolated DNA molecules (e.g., targeting vectors) for integrating a polynucleotide sequence encoding a transactivator of the invention at a predetermined location within a second target DNA molecule by homologous recombination are also disclosed. Transgenic mice having the DNA molecules of the invention integrated at a predetermined location in a chromosome by homologous recombination are also encompassed by the invention. Methods to regulate the expression of a tet operator linked-gene of interest by administering tetracycline or a tetracycline analogue to a mouse of the invention are also disclosed. The regulatory system of the invention allows for conditional inactivation or modulation of expression of a gene of interest in a host cell or mouse.

摘要翻译： 携带两个转基因的转基因小鼠，首先编码包含直接或间接在真核细胞中活化的tet阻遏物和多肽的反式激活因子融合蛋白，第二编码包含与可操作地连接至少一个tet操纵子的最小启动子可操作地连接的基因 序列。 还公开了通过同源重组将编码本发明的反式激活物的多核苷酸序列整合到第二靶DNA分子内的预定位置的分离的DNA分子（例如靶向载体）。 通过同源重组将具有整合在染色体中预定位置的本发明的DNA分子的转基因小鼠也包括在本发明中。 还公开了通过向本发明的小鼠施用四环素或四环素类似物来调节目的基因的表达的方法。 本发明的调节系统允许在宿主细胞或小鼠中条件性失活或调控感兴趣的基因的表达。

公开(公告)号：US5866755A

公开(公告)日：1999-02-02

申请号：US486814

申请日：1995-06-07

申请人： Hermann Bujard ,  Manfred Gossen

发明人： Hermann Bujard ,  Manfred Gossen

IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C07K14/035 ,  C07K14/195 ,  C07K14/245 ,  C07K19/00 ,  C12N1/15 ,  C12N1/19 ,  C12N5/10 ,  C12N15/09 ,  C12N15/11 ,  C12N15/12 ,  C12N15/62 ,  C12N15/63 ,  C12N15/67 ,  C12N15/85 ,  C12N5/00 ,  C12N15/00

CPC分类号： A01K67/0275 ,  C07K14/005 ,  C07K14/245 ,  C12N15/62 ,  C12N15/63 ,  C12N15/635 ,  C12N15/67 ,  C12N15/85 ,  C12N15/8509 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2217/20 ,  A01K2227/105 ,  A01K2267/0393 ,  A61K48/00 ,  C07K2319/00 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2710/16622 ,  C12N2830/003 ,  C12N2830/005 ,  C12N2830/006 ,  C12N2830/32

摘要： Transgenic animals carrying a transgene comprising a nucleic acid molecule encoding protein 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 and a second polypeptide which directly or indirectly inhibits transcription in eukaryotic cells. In various embodiment, the first polypeptide 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). In a preferred embodiment, the transgene encoding the transcriptional inhibitor fusion protein is integrated at a predetermined location within the chromosome of the transgenic animal.

摘要翻译： 公开了携带转基因的转基因，其包含编码可用于以高度受控的方式调控真核细胞和生物体中的基因表达的蛋白质的核酸分子。 在本发明的调节系统中，由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操纵子序列的第一多肽和直接或间接抑制转录的第二多肽 真核细胞。 在多种实施方案中，第一多肽与tet操纵子序列结合：（i）在不存在但不存在四环素（或其类似物）的情况下，或（ii）在存在但不存在四环素（或其类似物）的情况下 ）。 在优选的实施方案中，编码转录抑制剂融合蛋白​​的转基因整合在转基因动物的染色体内的预定位置。

公开(公告)号：US5859310A

公开(公告)日：1999-01-12

申请号：US481970

申请日：1995-06-07

申请人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

发明人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

IPC分类号： A01K67/027 ,  A61K48/00 ,  C12N5/08 ,  C12N15/62 ,  C12N15/63 ,  C12N15/85 ,  C12N5/00 ,  C12N15/00 ,  C12N15/09

CPC分类号： C12N15/62 ,  A01K67/0275 ,  C12N15/635 ,  C12N15/85 ,  A01K2217/05 ,  A01K2217/075 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2830/006

摘要： Transgenic mice carrying two transgenes, the first coding for a transactivator fusion protein comprising a tet repressor and a polypeptide which directly or indirectly activates transcription of a tet operator-linked gene in eucaryotic cells, and the second comprising a gene operably linked to a minimal promotor operably linked to at least one tet operator sequence, are disclosed. Isolated DNA molecules (e.g., targeting vectors) for integrating a polynucleotide sequence encoding a transactivator of the invention at a predetermined location within a second target DNA molecule by homologous recombination are also disclosed. Transgenic mice having the DNA molecules of the invention integrated at a predetermined location in a chromosome by homologous recombination are also encompassed by the invention. Methods to regulate the expression of a tet operator linked-gene of interest by administering tetracycline or a tetracycline analogue to a mouse of the invention are also disclosed. The regulatory system of the invention allows for conditional inactivation or modulation of expression of a gene of interest in a host cell or mouse.

摘要翻译： 携带两个转基因的转基因小鼠，首先编码包含tet阻遏物的反式激活因子融合蛋白，以及直接或间接激活真核细胞中与tet操纵子连接的基因的转录的多肽，第二编码包含可操作地连接到最小启动子的基因 可操作地连接至至少一个tet操纵子序列。 还公开了通过同源重组将编码本发明的反式激活物的多核苷酸序列整合到第二靶DNA分子内的预定位置的分离的DNA分子（例如靶向载体）。 通过同源重组将具有整合在染色体中预定位置的本发明的DNA分子的转基因小鼠也包括在本发明中。 还公开了通过向本发明的小鼠施用四环素或四环素类似物来调节目的基因的表达的方法。 本发明的调节系统允许在宿主细胞或小鼠中条件性失活或调控感兴趣的基因的表达。

公开(公告)号：US5464758A

公开(公告)日：1995-11-07

申请号：US76726

申请日：1993-06-14

申请人： Manfred Gossen ,  Hermann Bujard

发明人： Manfred Gossen ,  Hermann Bujard

IPC分类号： C07K14/035 ,  C12N15/63 ,  C12N15/85 ,  C12N15/09 ,  C12N5/10 ,  C12N15/11 ,  C12N15/62

CPC分类号： C12N15/85 ,  C07K14/005 ,  C12N15/635 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2710/16622 ,  C12N2830/006

摘要： A first polynucleotide molecule coding for a transactivator fusion protein comprising the tet repressor and a protein capable of activating transcription in eucaryotes. A second polynucleotide molecule coding for a protein, wherein said polynucleotide is operably linked to a minimal promoter operably linked to at least one tet operator sequence is also disclosed. A method to regulate the expression of a protein coded for by a polynucleotide, by cultivating the eucaryotic cell of the invention in a medium comprising tetracycline or a tetracycline analogue is also disclosed. Kits containing the polynucleotide molecules are also disclosed.

摘要翻译： 编码包含tet阻遏物和能够在真核生物中激活转录的蛋白质的反式激活因子融合蛋白的第一多核苷酸分子。 还公开了编码蛋白质的第二多核苷酸分子，其中所述多核苷酸可操作地连接到可操作地连接至至少一种tet操纵子序列的最小启动子。 还公开了通过在包含四环素或四环素类似物的培养基中培养本发明的真核细胞来调节多核苷酸编码的蛋白质的表达的方法。 还公开了含有多核苷酸分子的试剂盒。