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1. 发明申请

WO2006084695A3 TETRACYCLINE-CONTROLLED INSERTION ELEMENT FOR SYSTEMATIC MUTAGENESIS 审中-公开
标题翻译：用于系统MUTAGENESIS的TETRACYCLINE控制插入元件
公开(公告)号：WO2006084695A3
公开(公告)日：2006-11-02
申请号：PCT/EP2006001163
申请日：2006-02-09
申请人： UNIV FRIEDRICH ALEXANDER ER , HILLEN WOLFGANG , BERTRAM RALPH , KOESTNER MARTIN
发明人： HILLEN WOLFGANG , BERTRAM RALPH , KOESTNER MARTIN
IPC分类号： C12N15/82 , C12N15/71 , C12N15/74 , C12N15/85
CPC分类号： C12N15/8216 , C12N15/635 , C12N2800/30 , C12N2800/90 , C12N2820/55 , C12N2830/003
摘要： The present invention relates to an insertion element, wherein said insertion element is a recombinant nucleic acid molecule which comprises, in 5' ? 3' arrangement: (i) a first recombines binding site; (ii) a resistance gene, operatively linked to a promoter and, optionally, to transcription control sequences; (iii) a tetracycline-controlled promoter; and (iv) a second recombines binding site, wherein said tetracycline-controlled promoter is oriented to allow transcription in direction of the second recombines binding site. The present invention also relates to a method for the generation of a conditional knock-out mutant of a cell, comprising the steps of: (a) introducing a nucleic acid molecule comprising the insertion element of the present invention into a cell, wherein the insertion element is transfected into the cell concomitant with purified recombines; (b) integrating the insertion element into the cellular genome; and (c) selecting cells for resistance against a substrate for which the resistance gene of the insertion element confers resistance; wherein prior to or concomitant with step (a) an expression construct for a tetracycline-sensitive (poly)peptide is introduced into the cell, said (poly)peptide being capable of binding to the tetracycline-controlled promoter of the insertion element and of modulating its expression. Finally, the present invention relates to a cell containing the insertion element of the present invention, to a method for identifying genetic determinants of pathogen survival and to a method for the generation of a transgenic animal and plant.
摘要翻译：本发明涉及插入元件，其中所述插入元件是重组核酸分子，其包含在5' 3'排列：（i）首先重组结合位点; （ii）可操作地连接到启动子和任选地转录控制序列的抗性基因; （iii）四环素控制的启动子; 和（iv）第二重组结合位点，其中所述四环素受控启动子被取向以允许在第二重组结合位点的方向上转录。本发明还涉及一种用于产生细胞的条件敲除突变体的方法，包括以下步骤：（a）将包含本发明的插入元件的核酸分子导入细胞中，其中插入元素被转染到细胞中，伴随着纯化的重组; （b）将插入元件整合到细胞基因组中; 和（c）选择针对插入元件的抗性基因赋予抗性的底物的抗性的细胞; 其中在步骤（a）之前或伴随着步骤（a），将四环素敏感（多）肽的表达构建体引入细胞中，所述（多）肽能够结合插入元件的四环素受控启动子并调节其表达。最后，本发明涉及含有本发明的插入元件的细胞，鉴定鉴定病原体存活的遗传决定因素的方法以及产生转基因动物和植物的方法。

2. 发明申请

WO0075362A2 METHOD FOR IDENTIFYING NOVEL TRANSCRIPTIONAL REGULATORY PROTEINS 审中-公开
标题翻译：鉴定新型转录调节蛋白的方法
公开(公告)号：WO0075362A2
公开(公告)日：2000-12-14
申请号：PCT/IB0000859
申请日：2000-06-05
申请人： HILLEN WOLFGANG
发明人： HILLEN WOLFGANG
IPC分类号： C12N15/63 , C12Q1/68 , C12Q1/00
CPC分类号： C12Q1/6897 , C12N15/635
摘要： A method for identifying transcriptional regulatory proteins that modulate the transcription of a gene of interest is disclosed. In one embodiment, the method comprises introducing into a cell a nucleic acid molecule comprising three central components: 1) a polynucleotide ( e.g. , DNA) encoding a transcriptional regulatory protein; 2) an indicator gene which is responsive to, e.g. , under the transcriptional control of, the gene regulatory sequences of a gene of interest which bind to the transcriptional regulatory protein; and 3) a selectable marker gene. The transcriptional regulatory protein binds to the gene regulatory sequences and either activates or inhibits transcription. A protein is identified as a modulator of the transcription of the gene of interest by detecting a signal generated by the indicator gene.
摘要翻译：公开了一种用于鉴定调节感兴趣的基因的转录的转录调节蛋白的方法。在一个实施方案中，该方法包括向细胞中引入包含三个中心组分的核酸分子：1）编码转录调节蛋白的多核苷酸（例如，DNA）; 2）在与转录调节蛋白结合的感兴趣基因的基因调控序列的转录控制下，例如<！ - SIPO - >所响应的指示基因; 和3）选择标记基因。转录调节蛋白结合基因调控序列，并激活或抑制转录。通过检测由指示基因产生的信号，将蛋白质鉴定为感兴趣的基因的转录的调节剂。

3. 发明申请

WO0075347A3 TET REPRESSOR-BASED TRANSCRIPTIONAL REGULATORY PROTEINS 审中-公开
标题翻译：基于代谢基因的转录调节蛋白
公开(公告)号：WO0075347A3
公开(公告)日：2002-01-10
申请号：PCT/IB0000886
申请日：2000-06-05
申请人： HILLEN WOLFGANG , BUJARD HERMANN
发明人： HILLEN WOLFGANG , BUJARD HERMANN
IPC分类号： A01K67/027 , A61K31/65 , A61K35/74 , A61K35/76 , A61K48/00 , A61P31/12 , A61P35/00 , C07K14/47 , C07K19/00 , C12N1/19 , C12N5/10 , C12N15/09 , C12N15/63 , C12N15/85 , C12P21/02 , C12N15/62 , C07K14/16 , C12N15/00
CPC分类号： C12N15/635 , C07K2319/00 , C12N15/85 , C12N2800/108 , C12N2830/003 , C12N2830/006
摘要： The present invention provides a panel of transcriptional activator fusion proteins which comprises both tetracycline controlled transactivator proteins and reverse tetracycline transactivator proteins. These transactivators have novel phenotypes such as alterred basal transcriptional activity in the absence of doxycycline, altered induced transcriptional activity in the presence of doxycycline, or differential induction by tetracycline and analogs of tetracycline.
摘要翻译：本发明提供了一组转录激活物融合蛋白，其包含四环素控制的反式激活蛋白和反向四环素反式激活蛋白。这些反式激活剂具有新型表型，例如在多西环素不存在时改变的基础转录活性，在多西环素存在下改变的诱导转录活性，或四环素和四环素类似物的差异诱导。

4. 发明申请

WO2005123923A3 INDUCER SPECIFIC TETRACYCLINE REPRESSOR PROTEINS AND METHODS OF USE THEREOF 审中-公开
标题翻译：电感特异性四聚体代谢蛋白及其使用方法
公开(公告)号：WO2005123923A3
公开(公告)日：2006-06-15
申请号：PCT/EP2005006551
申请日：2005-06-17
申请人： HILLEN WOLFGANG , HENSSLER EVA
发明人： HILLEN WOLFGANG , HENSSLER EVA
IPC分类号： C12N15/63 , A01K67/027 , A61K38/16 , A61K39/40 , A61K48/00 , C07K14/195 , C07K16/12 , C12N5/10
CPC分类号： C12N15/635
摘要： The present invention relates to polynucleotides encoding inducer-specific tetracycline repressor proteins which are specifically induced by a tetracycline analog lacking a 4-dimethylamino grouping, vectors or host cells comprising said polynucleotides, or polypeptides being encoded thereby. Moreover, the present invention encompasses antibodies against the polypeptides, non-human transgenic animals comprising the polynucleotides or vectors or pharmaceutical compositions or uses. Finally, the present to invention pertains to methods for producing polypeptides or polynucleotides or for regulating transcription of a tet-operator-linked gene.
摘要翻译：本发明涉及编码由缺少4-二甲基氨基的四环素类似物，包含所述多核苷酸的载体或宿主细胞或由其编码的多肽特异性诱导的诱导物特异性四环素阻遏蛋白的多核苷酸。此外，本发明包括针对多肽的抗体，包含多核苷酸或载体或药物组合物或用途的非人转基因动物。最后，本发明涉及生产多肽或多核苷酸的方法或用于调节与tet操纵子连接的基因的转录。

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