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公开(公告)号：US20110014655A1
公开(公告)日：2011-01-20
申请号：US12807133
申请日：2010-08-26
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。
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公开(公告)号：US08039230B2
公开(公告)日：2011-10-18
申请号：US12226706
申请日：2007-04-24
申请人： Arie Pieter Otte , Henricus J. M. van Blokland , Theodorus H. J. Kwaks , Richard G. A. B. Sewalt
发明人： Arie Pieter Otte , Henricus J. M. van Blokland , Theodorus H. J. Kwaks , Richard G. A. B. Sewalt
CPC分类号： C12N15/67
摘要： The invention provides a DNA molecule comprising an open reading frame sequence that encodes a selectable marker polypeptide, wherein said DNA molecule in the coding strand comprises a translation start sequence for the selectable marker polypeptide having a GTG startcodon or a TTG startcodon, and wherein the open reading frame sequence that encodes the selectable marker protein has been mutated to replace at least half of its CpG dinucleotides as compared to the native open reading frame sequence that encodes the selectable marker protein.
摘要翻译： 本发明提供了包含编码选择性标记多肽的开放阅读框序列的DNA分子，其中编码链中的所述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，并且其中所述开放 与编码可选择标记蛋白的天然开放阅读框序列相比，编码可选择标记蛋白的阅读框序列已被突变以替代其至少一半的CpG二核苷酸。
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公开(公告)号：US08999667B2
公开(公告)日：2015-04-07
申请号：US11899505
申请日：2007-09-06
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。
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公开(公告)号：US08771984B2
公开(公告)日：2014-07-08
申请号：US12807133
申请日：2010-08-26
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。
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