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    • 10. 发明申请
    • NOVEL 88 PHAGE VECTORS
    • 88号新闻稿
    • WO03093471A8
    • 2004-07-22
    • PCT/US0214971
    • 2002-04-29
    • ALEXION PHARMA INC
    • BOWDISH KATHERINE SBARBAS-FREDERICKSON SHANA
    • C07H21/04C12N15/00C12N15/09C12N15/10C40B40/02C12P19/34
    • C40B40/02C07H21/04C12N15/1037
    • A phage genome is engineered to include a novel restriction site at one of two different positions. In a first embodiment, a restriction site is inserted into the phage genome between the end of gene IV and the MOS hairpin which serves as a phage packaging signal for newly synthesized single strands of phage DNA. In a second embodiment, a restriction site is inserted into the phage genome after the MOS hairpin and prior to the minus strand origin. Once the phage genome is modified to contain the new restriction site, the vector can be engineered to be a "88" vector by inserting at the new restriction site a nucleotide sequence encoding at least a functional domain of pVIII and at least a first cloning site for receiving a gene encoding a polypeptide to be displayed and, optionally a second cloning site for receiving to be displayed. In particularly useful embodiments, the novel vectors are engineered to produce phage particles that display antibodies.
    • 噬菌体基因组被设计为在两个不同位置之一包括新的限制性位点。 在第一实施方案中,将限制性位点插入到基因IV的末端和用作新合成的噬菌体DNA单链的噬菌体包装信号的MOS发夹之间的噬菌体基因组中。 在第二个实施方案中,将限制性位点插入到MOS发夹后并且在负链起始之前的噬菌体基因组中。 一旦噬菌体基因组被修饰以含有新的限制性位点,可以将该载体设计成“88”载体,通过在新的限制性位点插入编码至少pVIII的功能结构域的核苷酸序列和至少第一个克隆位点 用于接收编码要显示的多肽的基因,以及任选的用于接收待显示的第二克隆位点。 在特别有用的实施方案中,新载体被工程化以产生显示抗体的噬菌体颗粒。