公开(公告)号：US07192741B2

公开(公告)日：2007-03-20

申请号：US10190312

申请日：2002-07-05

申请人： Arie Peter Otte ,  Arthur Leo Kruckeberg

发明人： Arie Peter Otte ,  Arthur Leo Kruckeberg

IPC分类号： C12N5/10 ,  C12N5/18 ,  C12N15/00 ,  C12P21/00 ,  C07H21/04

CPC分类号： C12Q1/6897 ,  C07K14/4702 ,  C07K14/4705 ,  C12N15/1034 ,  C12N15/635 ,  C12N15/8216 ,  C12N15/8218

摘要： The invention is concerned with the systematic elucidation and identification of regulatory sequences. The invention provides among others screenings and detection methods with which regulatory sequences can be identified. The invention further provides regulatory sequences and use thereof in various fields such as, but not limited to protein production, diagnostics, transgenic plants and animals, and the therapeutic field.

公开(公告)号：US20070026498A1

公开(公告)日：2007-02-01

申请号：US11580605

申请日：2006-10-13

申请人： Arie Otte ,  Arthur Kruckeberg

发明人： Arie Otte ,  Arthur Kruckeberg

IPC分类号： C12N15/86 ,  C07H21/04 ,  C12P21/06 ,  C07K14/47 ,  C12N5/06

CPC分类号： C12Q1/6897 ,  C07K14/4702 ,  C07K14/4705 ,  C12N15/1034 ,  C12N15/635 ,  C12N15/8216 ,  C12N15/8218

摘要： The invention is concerned with the systematic elucidation and identification of regulatory sequences. The invention provides among others screenings and detection methods with which regulatory sequences can be identified. The invention further provides regulatory sequences and use thereof in various fields such as, but not limited to, protein production, diagnostics, transgenic plants and animals, and the therapeutic field.

公开(公告)号：US06872524B1

公开(公告)日：2005-03-29

申请号：US09762916

申请日：1999-08-16

申请人： Arie Otte

发明人： Arie Otte

IPC分类号： C12N15/09 ,  C12Q1/02 ,  C12Q1/68

CPC分类号： C12Q1/6897

摘要： The invention relates to a method of detecting a DNA sequence which at least partially contributes to promote the stable expression of a gene. To this end the DNA fragment to be examined is cloned in a vector between i) a DNA sequence involved in the induction of gene transcription repressing chromatin, and ii) a reporter gene. The invention also relates to a DNA sequence to be detected by means of the invention, and the application of a stable expression-enhancing DNA sequence for the stable expression of a gene.

摘要翻译： 本发明涉及检测至少部分促进基因稳定表达的DNA序列的方法。 为此，将待检查的DNA片段克隆在载体中，i）涉及诱导基因转录抑制染色质的DNA序列，和ii）报告基因。 本发明还涉及通过本发明检测的DNA序列和用于稳定表达基因的稳定表达增强DNA序列的应用。

公开(公告)号：US09228004B2

公开(公告)日：2016-01-05

申请号：US14081938

申请日：2013-11-15

申请人： ChromaGenics B.V.

发明人： Arie Pieter Otte ,  Henricus J.M. van Blokland ,  Theodorus Hendrikus Jacobus Kwaks ,  Richard George Antonius Bernardus Sewalt

IPC分类号： C12P21/06 ,  C07K14/505 ,  C12N15/85 ,  C12N9/12 ,  C12P21/04 ,  C12N1/00 ,  C12N5/02 ,  C12N15/00 ,  C12N15/74 ,  C12N15/82 ,  C12N15/86 ,  C07H21/00 ,  C07H21/04

CPC分类号： C07K14/505 ,  C12N9/1205 ,  C12N15/67 ,  C12N15/85 ,  C12N2830/46 ,  C12N2840/20 ,  C12N2840/203 ,  C12N2840/206 ,  C12N2840/50 ,  C12Y207/01095

摘要： Described is a DNA molecule comprising an open reading frame sequence encoding a selectable marker polypeptide, wherein the DNA molecule in the coding strand comprises a translation start sequence for the selectable marker polypeptide having a GTG or TTG start codon, and wherein the ORF sequence that encodes the selectable marker protein has been mutated to replace at least half of its CpG dinucleotides as compared to the native ORF sequence that encodes the selectable marker protein. Further provided are such DNA molecules wherein the ORF sequence that encodes a selectable marker polypeptide is part of a multicistronic transcription unit that further comprises an open reading frame sequence encoding a polypeptide of interest. Also described are methods for obtaining host cells expressing a polypeptide of interest, wherein the host cells comprise the DNA molecules described herein. Further provided is the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules described herein.

摘要翻译： 描述了包含编码可选择标记多肽的开放阅读框序列的DNA分子，其中编码链中的DNA分子包含具有GTG或TTG起始密码子的可选择标记多肽的翻译起始序列，其中编码 与编码选择性标记蛋白的天然ORF序列相比，可选择标记蛋白已被突变以替代其至少一半的CpG二核苷酸。 还提供了这样的DNA分子，其中编码可选择标记多肽的ORF序列是多顺反子转录单位的一部分，其进一步包含编码目标多肽的开放阅读框序列。 还描述了获得表达目的多肽的宿主细胞的方法，其中宿主细胞包含本文所述的DNA分子。 还提供感兴趣的多肽的生产，包括培养包含本文所述的DNA分子的宿主细胞。

公开(公告)号：US08999667B2

公开(公告)日：2015-04-07

申请号：US11899505

申请日：2007-09-06

申请人： Arie P. Otte ,  Theodorus H. J. Kwaks ,  Henricus J. M. van Blokland ,  Richard G. A. B. Sewalt

发明人： Arie P. Otte ,  Theodorus H. J. Kwaks ,  Henricus J. M. van Blokland ,  Richard G. A. B. Sewalt

IPC分类号： C12N15/63 ,  C12N15/65 ,  C12N15/67 ,  C12N15/79 ,  C12N15/85

CPC分类号： C12N15/85 ,  C12N2830/46 ,  C12N2840/203

摘要： The invention provides a DNA molecule comprising a multicistronic transcription unit coding for i) a polypeptide of interest, and for ii) a selectable marker polypeptide functional in a eukaryotic host cell, wherein the polypeptide of interest has a translation initiation sequence separate from that of the selectable marker polypeptide, and wherein the coding sequence for the polypeptide of interest is upstream from the coding sequence for the selectable marker polypeptide in the multicistronic transcription unit, and wherein an internal ribosome entry site is present downstream from the coding sequence for the polypeptide of interest and upstream from the coding sequence for the selectable marker polypeptide, and wherein the nucleic acid sequence coding for the selectable marker polypeptide in the coding strand comprises a GTG or a TTG start codon. Also provided are methods for obtaining host cells expressing a polypeptide of interest, such host cells comprising DNA molecules of the invention. Further provided is the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules of the invention.

摘要翻译： 本发明提供了一种DNA分子，其包含编码i）感兴趣多肽的多顺反子转录单位，以及ii）在真核宿主细胞中有功能的选择性标记多肽，其中所述目标多肽具有与 选择性标记多肽，并且其中感兴趣的多肽的编码序列在多顺反子转录单位中的选择性标记多肽的编码序列的上游，并且其中内部核糖体进入位点存在于感兴趣的多肽的编码序列的下游 并且编码选择性标记多肽的编码序列的上游，并且其中编码链中编码选择性标记多肽的核酸序列包含GTG或TTG起始密码子。 还提供了获得表达目的多肽的宿主细胞的方法，所述宿主细胞包含本发明的DNA分子。 进一步提供感兴趣的多肽的生产，包括培养包含本发明的DNA分子的宿主细胞。

公开(公告)号：US08771984B2

公开(公告)日：2014-07-08

申请号：US12807133

申请日：2010-08-26

申请人： Arie P. Otte ,  Henricus J. M. van Blokland ,  Theodorus H. J. Kwaks ,  Richard G. A. B. Sewalt

发明人： Arie P. Otte ,  Henricus J. M. van Blokland ,  Theodorus H. J. Kwaks ,  Richard G. A. B. Sewalt

IPC分类号： C12N15/00 ,  C12N15/09 ,  C12N15/63 ,  C07K14/00

CPC分类号： C07K16/00 ,  C07K14/00 ,  C07K16/30

摘要： The invention provides a DNA molecule comprising a multicistronic transcription unit coding for i) a selectable marker polypeptide functional in a eukaryotic host cell, and for ii) a polypeptide of interest, the polypeptide of interest having a translation initiation sequence separate from that of the selectable marker polypeptide, characterized in that the coding sequence for the polypeptide of interest is downstream from the coding sequence for the selectable marker in the multicistronic transcription unit, and the nucleic acid sequence coding for the selectable marker polypeptide comprises a mutation that decreases the translation efficiency of the selectable marker in a eukaryotic host cell. The invention also provides methods for obtaining host cells expressing a polypeptide of interest, the host cells comprising the DNA molecules of the invention. The invention further provides the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules according to the invention.

摘要翻译： 本发明提供了一种DNA分子，其包含编码i）在真核宿主细胞中有功能的选择性标记多肽，以及ii）感兴趣的多肽的多顺反子转录单位，所述感兴趣的多肽具有与可选择的不同的翻译起始序列 标记多肽，其特征在于感兴趣的多肽的编码序列在多顺反子转录单位中的选择性标记的编码序列的下游，并且编码选择标记多肽的核酸序列包含降低翻译效率的突变 在真核宿主细胞中的选择性标记。 本发明还提供了获得表达目的多肽的宿主细胞的方法，宿主细胞包含本发明的DNA分子。 本发明进一步提供感兴趣的多肽的生产，包括培养包含根据本发明的DNA分子的宿主细胞。

公开(公告)号：US20140106401A1

公开(公告)日：2014-04-17

申请号：US14081938

申请日：2013-11-15

申请人： ChromaGenics B.V.

发明人： Arie Pieter Otte ,  Henricus J.M. van Blokland ,  Theodorus Hendrikus Jacobus Kwaks ,  Richard George Antonius Bernardus Sewalt

IPC分类号： C07K14/505

CPC分类号： C07K14/505 ,  C12N9/1205 ,  C12N15/67 ,  C12N15/85 ,  C12N2830/46 ,  C12N2840/20 ,  C12N2840/203 ,  C12N2840/206 ,  C12N2840/50 ,  C12Y207/01095

摘要： Described is a DNA molecule comprising an open reading frame sequence encoding a selectable marker polypeptide, wherein the DNA molecule in the coding strand comprises a translation start sequence for the selectable marker polypeptide having a GTG or TTG start codon, and wherein the ORF sequence that encodes the selectable marker protein has been mutated to replace at least half of its CpG dinucleotides as compared to the native ORF sequence that encodes the selectable marker protein. Further provided are such DNA molecules wherein the ORF sequence that encodes a selectable marker polypeptide is part of a multicistronic transcription unit that further comprises an open reading frame sequence encoding a polypeptide of interest. Also described are methods for obtaining host cells expressing a polypeptide of interest, wherein the host cells comprise the DNA molecules described herein. Further provided is the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules described herein.

摘要翻译： 描述了包含编码可选择标记多肽的开放阅读框序列的DNA分子，其中编码链中的DNA分子包含具有GTG或TTG起始密码子的选择性标记多肽的翻译起始序列，其中编码 与编码选择性标记蛋白的天然ORF序列相比，可选择标记蛋白已被突变以替代其至少一半的CpG二核苷酸。 还提供了这样的DNA分子，其中编码可选择标记多肽的ORF序列是多顺反子转录单位的一部分，其进一步包含编码目标多肽的开放阅读框序列。 还描述了获得表达目的多肽的宿主细胞的方法，其中宿主细胞包含本文所述的DNA分子。 还提供感兴趣的多肽的生产，包括培养包含本文所述的DNA分子的宿主细胞。

公开(公告)号：US08263393B2

公开(公告)日：2012-09-11

申请号：US11156910

申请日：2005-06-20

申请人： Arie Pieter Otte ,  Theodorus Hendrikus Jacobus Kwaks ,  Richard George Antonius Bernardus Sewalt

发明人： Arie Pieter Otte ,  Theodorus Hendrikus Jacobus Kwaks ,  Richard George Antonius Bernardus Sewalt

IPC分类号： C12N15/00 ,  C12N15/63 ,  C07H21/04

CPC分类号： C12N15/635 ,  C12N15/63 ,  C12N15/85 ,  C12N2830/48 ,  C12N2840/203 ,  C12N2840/206

摘要： Described are means and methods for providing a cell with a protein expression unit, the method comprising providing a nucleic acid sequence comprising the unit with a nucleic acid sequence encoding a binding site for a member of a chromatin modification system for rendering chromatin more accessible for transcription (opener), wherein the opener is present in the cell. Preferred openers comprise histone modification proteins, chromatin remodeling proteins and trithorax group proteins or equivalents. The cells thus generated and nucleic acid sequences encoding such openers are provided. Openers are preferred in the context of STAR and TRAP sequences.

摘要翻译： 描述了用于向细胞提供蛋白质表达单元的方法和方法，所述方法包括提供包含该单元的核酸序列，该核酸序列编码编码染色质修饰系统成员的结合位点的核酸序列，以使得染色质更易于转录 （开环剂），其中开启剂存在于电池中。 优选的开放剂包括组蛋白修饰蛋白，染色质重塑蛋白和trithorax组蛋白或等同物。 提供由此产生的细胞和编码这种开放子的核酸序列。 在STAR和TRAP序列的背景下，开放者是首选的。

公开(公告)号：US20110300580A1

公开(公告)日：2011-12-08

申请号：US13135966

申请日：2011-07-18

申请人： Arie Pieter Otte ,  Henricus Johannes Maria Van Blokland ,  Theodorus Hendrikus Jacobus Kwaks ,  Richard George Antonius Bernardus Sewalt

发明人： Arie Pieter Otte ,  Henricus Johannes Maria Van Blokland ,  Theodorus Hendrikus Jacobus Kwaks ,  Richard George Antonius Bernardus Sewalt

IPC分类号： C12P21/02 ,  C12N15/85 ,  C12N15/63 ,  C12N1/00 ,  C12N1/21 ,  C07H21/04 ,  C12N5/10

CPC分类号： C07K14/505 ,  C12N9/1205 ,  C12N15/67 ,  C12N15/85 ,  C12N2830/46 ,  C12N2840/20 ,  C12N2840/203 ,  C12N2840/206 ,  C12N2840/50 ,  C12Y207/01095

摘要： Described is a DNA molecule comprising an open reading frame sequence that encodes a selectable marker polypeptide, wherein the DNA molecule in the coding strand comprises a translation start sequence for the selectable marker polypeptide having a GTG start codon or a TTG start codon, and wherein the ORF sequence that encodes the selectable marker protein has been mutated to replace at least half of its CpG dinucleotides as compared to the native ORF sequence that encodes the selectable marker protein. Further provided are such DNA molecules wherein the ORF sequence that encodes a selectable marker polypeptide is part of a multicistronic transcription unit that further comprises an open reading frame sequence encoding a polypeptide of interest. Also described are methods for obtaining host cells expressing a polypeptide of interest, wherein the host cells comprise the DNA molecules described herein. Further provided is the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules described herein.

摘要翻译： 描述了包含编码可选择标记多肽的开放阅读框序列的DNA分子，其中编码链中的DNA分子包含具有GTG起始密码子或TTG起始密码子的可选择标记多肽的翻译起始序列，其中 与编码可选择标记蛋白的天然ORF序列相比，编码可选择标记蛋白的ORF序列已被突变以替代其至少一半的CpG二核苷酸。 还提供了这样的DNA分子，其中编码可选择标记多肽的ORF序列是多顺反子转录单位的一部分，其进一步包含编码目标多肽的开放阅读框序列。 还描述了获得表达目的多肽的宿主细胞的方法，其中宿主细胞包含本文所述的DNA分子。 还提供感兴趣的多肽的生产，包括培养包含本文所述的DNA分子的宿主细胞。

公开(公告)号：US07901908B2

公开(公告)日：2011-03-08

申请号：US11978134

申请日：2007-10-26

申请人： Arie Pieter Otte ,  Arthur Leo Kruckeberg ,  Richard George Antonius Bernardus Sewalt

发明人： Arie Pieter Otte ,  Arthur Leo Kruckeberg ,  Richard George Antonius Bernardus Sewalt

IPC分类号： C12P21/02

CPC分类号： C07K16/00 ,  C07K16/18 ,  C12N15/67 ,  C12N15/8218 ,  C12N15/822 ,  C12N15/8257

摘要： Described is the production of proteins in a host cell. More specifically, described are methods for improving expression of two or more proteins in a cell or host cell. The methods are suited for production of, for example, recombinant antibodies that can be used in pharmaceutical preparations or as diagnostic tools. In one embodiment, provided is a method for obtaining a cell that expresses two or more proteins comprising providing the cell with two or more protein expression units encoding two or more proteins, characterized in that at least two of the protein expression units comprise at least one STAR sequence.

摘要翻译： 描述了宿主细胞中蛋白质的产生。 更具体地，描述了用于改善细胞或宿主细胞中两种或更多种蛋白质的表达的方法。 该方法适用于生产例如可用于药物制剂或诊断工具的重组抗体。 在一个实施方案中，提供了获得表达两种或更多种蛋白质的细胞的方法，包括向细胞提供两种或更多种编码两种或更多种蛋白质的蛋白质表达单位，其特征在于至少两种蛋白质表达单位包含至少一种 STAR序列。