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公开(公告)号：US09102971B2
公开(公告)日：2015-08-11
申请号：US13326873
申请日：2011-12-15
申请人： Megumi Kurokawa , Keina Yamaguchi , Risa Ogawa , Masayoshi Tsukahara , Koichi Kawakami , Yoko Hayashi
发明人： Megumi Kurokawa , Keina Yamaguchi , Risa Ogawa , Masayoshi Tsukahara , Koichi Kawakami , Yoko Hayashi
CPC分类号： C12P21/02 , C07K16/1018 , C07K16/241 , C07K16/2887 , C07K16/2896 , C07K2317/14 , C12N15/85 , C12N2800/107 , C12N2800/90
摘要： The present invention relates to a method for integrating a gene fragment inserted between a pair of transposon sequences into a chromosome of a mammalian cell, comprising introducing at least one expression vector which comprises a gene fragment comprising a DNA encoding a protein of interest and also comprises a pair of transposon sequences at both terminals of the gene fragment, into a suspension mammalian cell; and a method for producing the protein of interest comprising suspension-culturing a suspension mammalian cell which produces the protein of interest; and an a suspension mammalian cell which expresses the protein of interest.
摘要翻译： 本发明涉及将插入在一对转座序列之间的基因片段整合到哺乳动物细胞的染色体中的方法，包括引入至少一种表达载体，所述表达载体包含含有编码感兴趣的蛋白质的DNA的基因片段，并且还包含 在基因片段的两个末端的一对转座子序列转化成悬浮的哺乳动物细胞; 以及用于产生目的蛋白质的方法，包括悬浮培养产生目的蛋白质的悬浮哺乳动物细胞; 和表达目的蛋白质的悬浮哺乳动物细胞。
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公开(公告)号：US09034649B2
公开(公告)日：2015-05-19
申请号：US12813920
申请日：2010-06-11
CPC分类号： C12P21/00 , C07K16/1018 , C12N15/85 , C12N2015/8518 , C12N2800/107 , C12N2800/90 , C12N2840/20 , C12P21/02
摘要： This invention relates to a method for producing a protein of interest, comprising introducing a protein expression vector which comprises a gene fragment a gene fragment comprising a DNA encoding a protein of interest and a selectable marker gene and transposon sequences at both terminals of the gene fragment, into a suspension mammalian cell; integrating the gene fragment inserted between a pair of the transposon sequences, into a chromosome of the mammalian cell to obtain a mammalian cell capable of expressing the protein of interest; and suspension-culturing the mammalian cell; and a suspension mammalian cell capable of expressing the protein of interest.
摘要翻译： 本发明涉及一种生产目的蛋白质的方法，包括引入蛋白质表达载体，其包含基因片段，包含编码目标蛋白的DNA和选择性标记基因的基因片段，以及基因片段两端的转座子序列 ，进入悬浮哺乳动物细胞; 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，以获得能够表达目的蛋白质的哺乳动物细胞; 并悬浮培养哺乳动物细胞; 和能够表达目标蛋白质的悬浮哺乳动物细胞。
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公开(公告)号：US09410176B2
公开(公告)日：2016-08-09
申请号：US13994178
申请日：2011-12-14
CPC分类号： C12P21/00 , C07K16/00 , C07K16/2896 , C07K2317/14 , C12N15/85 , C12N2800/107 , C12N2800/90 , C12P21/02
摘要： This invention relates to a method for producing a protein of interest, comprising introducing an expression vector which comprises a gene fragment comprising a DNA encoding the protein of interest and a selectable marker gene and also comprises a pair of transposon sequences at both terminals of the gene fragment, into a suspension mammalian cell; integrating the gene fragment inserted between the pair of transposon sequences into a chromosome of the mammalian cell; obtaining a suspension mammalian cell producing the protein of interest; and suspension-culturing the suspension mammalian cell, and a suspension mammalian cell which expresses the protein of interest by the method.
摘要翻译： 本发明涉及一种生产目的蛋白质的方法，包括引入包含基因片段的表达载体，所述基因片段包含编码目的蛋白质的DNA和选择性标记基因，并且还包含在基因的两个末端的一对转座子序列 片段，成悬浮哺乳动物细胞; 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; 获得产生目的蛋白质的悬浮哺乳动物细胞; 悬浮培养悬浮哺乳动物细胞，以及通过该方法表达目的蛋白质的悬浮哺乳动物细胞。
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公开(公告)号：US20130273604A1
公开(公告)日：2013-10-17
申请号：US13994178
申请日：2011-12-14
IPC分类号： C12P21/00
CPC分类号： C12P21/00 , C07K16/00 , C07K16/2896 , C07K2317/14 , C12N15/85 , C12N2800/107 , C12N2800/90 , C12P21/02
摘要： This invention relates to a method for producing a protein of interest, comprising introducing an expression vector which comprises a gene fragment comprising a DNA encoding the protein of interest and a selectable marker gene and also comprises a pair of transposon sequences at both terminals of the gene fragment, into a suspension mammalian cell; integrating the gene fragment inserted between the pair of transposon sequences into a chromosome of the mammalian cell; obtaining a suspension mammalian cell producing the protein of interest; and suspension-culturing the suspension mammalian cell, and a suspension mammalian cell which expresses the protein of interest by the method.
摘要翻译： 本发明涉及一种生产目的蛋白质的方法，包括引入包含基因片段的表达载体，所述基因片段包含编码目的蛋白质的DNA和选择性标记基因，并且还包含在基因的两个末端的一对转座子序列 片段，成悬浮哺乳动物细胞; 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; 获得产生目的蛋白质的悬浮哺乳动物细胞; 悬浮培养悬浮哺乳动物细胞，以及通过该方法表达目的蛋白质的悬浮哺乳动物细胞。
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