公开(公告)号：US20060195935A1

公开(公告)日：2006-08-31

申请号：US11416490

申请日：2006-05-02

申请人： Arie Otte ,  Henricus Van Blokland ,  Theodorus Kwaks ,  Richard Bernardus Sewalt

发明人： Arie Otte ,  Henricus Van Blokland ,  Theodorus Kwaks ,  Richard Bernardus Sewalt

IPC分类号： A01H1/00 ,  C07H21/04 ,  C12N15/82 ,  C12N5/04

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： 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. The invention further provides such DNA molecules wherein the open reading frame 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. The invention also provides methods for obtaining host cells expressing a polypeptide of interest, said 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。 本发明还提供了这样的DNA分子，其中编码可选择标记多肽的开放阅读框序列是多顺反子转录单位的一部分，其进一步包含编码目的多肽的开放阅读框序列。 本发明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。 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：US20060003416A1

公开(公告)日：2006-01-05

申请号：US11156910

申请日：2005-06-20

申请人： Arie Otte ,  Theodorus Kwaks ,  Richard Bernardus Sewalt

发明人： Arie Otte ,  Theodorus Kwaks ,  Richard Bernardus Sewalt

IPC分类号： C12Q1/68 ,  C12P21/06 ,  C12N9/10 ,  C12N15/85

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。