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    • 3. 发明申请
    • Process for producing functional non-naturally occurring proteins, and method for site-specific modification and immobilization of the proteins
    • 用于产生功能非天然蛋白质的方法,以及用于位点特异性修饰和固定蛋白质的方法
    • US20090011459A1
    • 2009-01-08
    • US12073233
    • 2008-03-03
    • Daisuke KigaMasahiko Uchiyama
    • Daisuke KigaMasahiko Uchiyama
    • C12P21/02C07K1/00
    • C12P21/02C12N15/102
    • There is provided a process for industrial production of non-naturally occurring proteins composed of less than 20 amino acids, wherein the proteins retain their original functions while being capable of site-specific modification or immobilization, or having new functions not found in nature in addition to the original functions of the proteins.Specifically there are provided a process for producing a functional non-naturally occurring protein having a specific amino acid type(s) replaced with a natural amino acid(s) other than the amino acid type(s), the process comprising: a) matching a nucleic acid portion having a nucleotide sequence reflecting the genotype with a protein portion that is the translation product of the nucleic acid portion; b) selecting the matched molecule obtained in step a); c) introducing mutation into the nucleic acid portion of the matched molecule obtained in step b); d) amplifying the nucleic acid portion obtained in step c); e) providing the nucleic acid portion obtained in step d) to step a), to match the nucleic acid portion with a protein portion that is the translation product of the nucleic acid portion; and f) selecting the matched molecule obtained in step e), to produce a functional non-naturally occurring protein; and a method for site-specific modification and site-immobilization of a functional non-naturally occurring protein.
    • 提供了由少于20个氨基酸组成的非天然存在的蛋白质的工业生产方法,其中蛋白质保持其原始功能,同时能够进行位点特异性修饰或固定化,或具有另外在自然界中未发现的新功能 到蛋白质的原始功能。 具体地说,提供了一种制备具有被除了氨基酸类型之外的天然氨基酸替换的特定氨基酸类型的功能性非天然存在蛋白质的方法,该方法包括:a)匹配 具有反映基因型的核苷酸序列的核酸部分与作为核酸部分的翻译产物的蛋白质部分; b)选择在步骤a)中获得的匹配分子; c)将突变引入步骤b)中获得的匹配分子的核酸部分; d)扩增步骤c)中获得的核酸部分; e)提供步骤d)中获得的核酸部分至步骤a),以使核酸部分与作为核酸部分的翻译产物的蛋白质部分相匹配; 和f)选择步骤e)中获得的匹配分子,以产生功能非天然存在的蛋白质; 以及用于功能性非天然存在蛋白质的位点特异性修饰和位点固定的方法。