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    タンパク質の生産方法
    6.
    发明专利
    タンパク質の生産方法 有权
    生产蛋白质的方法

    公开(公告)号：JP2015226541A

    公开(公告)日：2015-12-17

    申请号：JP2015144268

    申请日：2015-07-21

    申请人： 大学共同利用機関法人情報・システム研究機構 ,  協和発酵キリン株式会社

    发明人： 川上 浩一 ,  山口 恵奈 ,  小川 梨沙 ,  塚原 正義

    IPC分类号： C12N5/10 ,  C12N15/09 ,  C12P21/00

    CPC分类号： C12P21/00 ,  C07K16/1018 ,  C12N15/85 ,  C12P21/02 ,  C12N2015/8518 ,  C12N2800/107 ,  C12N2800/90 ,  C12N2840/20

    摘要： 【課題】目的タンパク質をコードするDNAを哺乳動物細胞に導入して、目的タンパク質を生産する方法の提供。 【解決手段】目的タンパク質をコードするDNAおよび選択マーカー遺伝子を含む遺伝子断片の両端にトランスポゾン配列を含むタンパク質発現ベクターを浮遊性の哺乳動物細胞に導入し、一対のトランスポゾン配列の間に挿入された該遺伝子断片を該哺乳動物細胞の染色体に組み込んで該目的タンパク質を発現する哺乳動物細胞を得て、かつ該哺乳動物細胞を浮遊培養して該目的タンパク質を生産する方法。 【選択図】なし

    摘要翻译： 要解决的问题：提供通过将编码期望的蛋白质的DNA导入哺乳动物细胞来制备所需蛋白质的方法。解决方案：本发明提供了一种通过在两者中引入包含转座子序列的蛋白质表达载体来产生所需蛋白质的方法 包含编码所需蛋白质的DNA和选择标记基因的基因片段的末端插入到浮动的哺乳动物细胞中，将插入在一对转座子序列之间的基因片段整合到哺乳动物细胞的染色体中，以获得表达 所需蛋白质，并悬浮培养哺乳动物细胞。

    Expression of the endogenous gene by non-homologous recombination with the cell dna vector constructs
    9.
    发明专利
    Expression of the endogenous gene by non-homologous recombination with the cell dna vector constructs 有权

    公开(公告)号：JP5377806B2

    公开(公告)日：2013-12-25

    申请号：JP2000512942

    申请日：1998-09-25

    申请人： アサーシス,インコーポレイテッド

    发明人： ハリントン，ジョン・ジェイ

    IPC分类号： C12N15/09 ,  A61K48/00 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N15/67 ,  C12N15/85 ,  C12N15/90 ,  C12P21/00

    CPC分类号： C12N15/90 ,  A61K48/00 ,  C12N15/67 ,  C12N15/85 ,  C12N2840/20 ,  C12N2840/44

    摘要： The present invention relates generally to activating gene expression or causing over-expression of a gene by recombination methods in situ. The invention also relates generally to methods for expressing an endogenous gene in a cell at levels higher than those normally found in the cell. In one embodiment of the invention, expression of an endogenous gene is activated or increased following integration into the cell, by non-homologous or illegitimate recombination, of a regulatory sequence that activates expression of the gene. In another embodiment, the expression of the endogenous gene may be further increased by co-integration of one or more amplifiable markers, and selecting for increased copies of the one or more amplifiable markers located on the integrated vector. The invention also provides methods for the identification and expression of genes undiscoverable by current methods since no target sequence is necessary for integration. The invention also provides methods for isolation of nucleic acid molecules (particularly cDNA molecules) encoding transmembrane proteins, and for isolation of cells expressing such transmembrane proteins which may be heterologous transmembrane proteins. The invention also relates to isolated genes, gene products, nucleic acid molecules, and compositions comprising such genes, gene products and nucleic acid molecules, that may be used in a variety of therapeutic and diagnostic applications. Thus, by the present invention, endogenous genes, including those associated with human disease and development, may be activated and isolated without prior knowledge of the sequence, structure, function, or expression profile of the genes.