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    • 2. 发明申请
    • Method of sorting vesicle-entrapped, coupled nucleic acid-protein displays
    • 分离囊泡夹带的偶联核酸蛋白质显示器的方法
    • US20040265835A1
    • 2004-12-30
    • US10610024
    • 2003-06-30
    • David M. LemasterJames H. Jett
    • C12Q001/68C07H021/04C12P021/04C12N009/00C12N001/18
    • C40B40/02C07H21/04C12N15/1037
    • A method for identifying and sorting variants of a chosen enzyme is disclosed. Enzyme variants of a chosen enzyme are obtained and then linked to their corresponding genetic code through any of a family of suitable surface display methods. The enzyme variants, now displayed on the surface of a biological particle such as a phage, virus, yeast, or bacterium are then encapsulated in a vesicle containing an enzyme activity-sensitive assay reagent. The enzyme variant is thus exposed to the assay reagent, and displays a signal using the enzyme activity-sensitive assay reagent in a manner proportionate to the levels of activity of the enzyme, thus rendering the vesicles suitable for mechanical sorting based on these levels. The vesicles are then sorted using methods known in the art to isolate those variants exhibiting possibly beneficial variations in enzyme function.
    • 公开了用于鉴定和分选所选酶的变体的方法。 获得选择的酶的酶变体,然后通过任何一种合适的表面显示方法与其相应的遗传密码连接。 然后将现在显示在诸如噬菌体,病毒,酵母或细菌的生物颗粒的表面上的酶变体包封在含有酶活性敏感性测定试剂的囊泡中。 因此,酶变体暴露于测定试剂,并且以与酶的活性水平成比例的方式使用酶活性敏感的测定试剂显示信号,从而使得囊泡适合于基于这些水平的机械分选。 然后使用本领域已知的方法分选囊泡,以分离显示可能有益的酶功能变化的那些变体。