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    • 30. 发明专利
    • Improved mammal expression vector and use thereof
    • 改进的主动表达载体及其用途
    • JP2014147390A
    • 2014-08-21
    • JP2014058227
    • 2014-03-20
    • Abbvie Incアッヴィ・インコーポレイテッド
    • HSIEH CHUNG-MING
    • C12N15/09C12N5/10C12P21/02
    • C12P21/00C07K16/241C07K2317/21C07K2317/52
    • PROBLEM TO BE SOLVED: To provide a nucleic acid for a recombinant protein expression in mammalian cell culture.SOLUTION: Disclosed is an expression vector comprising (a) OriP origin of replication derived from Epstein-Barr virus (EBV); (b) a SV40 origin of replication; (c) an insertion site for inserting a gene of interest; (d) s cytomegalovirus (CMV) promoter operably linked to the insertion site, wherein the OriP origin of replication is bound by a trans-acting EBNA1 replication initiation factor which is not encoded by the expression vector, or the SV40 origin of replication is bound by a trans-acting SV40 T antigen which is not encoded by the expression vector. The episomal vectors promote high protein production in mammalian cells expressing the SV40 T Ag or Epstein-Barr virus nuclear antigen (e.g., COS7 or HEK293-6E cells).
    • 要解决的问题:提供哺乳动物细胞培养物中重组蛋白表达的核酸。解决方案:公开了包含(a)衍生自爱泼斯坦 - 巴尔病毒(EBV)的OriP复制起点的表达载体; (b)SV40复制起点; (c)用于插入感兴趣的基因的插入位点; (d)可操作地连接到插入位点的巨细胞病毒(CMV)启动子,其中所述的OriP复制起点由不由表达载体编码的反式作用EBNA1复制起始因子结合,或者SV40复制起点被结合 通过不由表达载体编码的反式作用SV40T抗原。 附加型载体促进表达SV40T Ag或爱泼斯坦 - 巴尔病毒核抗原(例如COS7或HEK293-6E细胞)的哺乳动物细胞中的高蛋白质产生。