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1. 发明申请

US20090011459A1 Process for producing functional non-naturally occurring proteins, and method for site-specific modification and immobilization of the proteins 有权
标题翻译：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
公开(公告)号：US20090011459A1
公开(公告)日：2009-01-08
申请号：US12073233
申请日：2008-03-03
申请人： Daisuke Kiga , Masahiko Uchiyama
发明人： Daisuke Kiga , Masahiko Uchiyama
IPC分类号： C12P21/02 , C07K1/00
CPC分类号： C12P21/02 , C12N15/102
摘要： 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.
摘要翻译：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。

2. 发明授权

US08227209B2 Process for producing functional non-naturally occurring proteins, and method for site-specific modification and immobilization of the proteins 有权
标题翻译：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
公开(公告)号：US08227209B2
公开(公告)日：2012-07-24
申请号：US12073233
申请日：2008-03-03
申请人： Daisuke Kiga , Masahiko Uchiyama
发明人： Daisuke Kiga , Masahiko Uchiyama
IPC分类号： C12N15/00 , C12N15/11
CPC分类号： C12P21/02 , C12N15/102
摘要： 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).
摘要翻译：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，提供了一种用氨基酸类型以外的天然氨基酸替代特定氨基酸类型的功能性非天然存在蛋白质的方法。

3. 发明申请

US20050084856A1 Tyrosyl-trna syntiiase variants 失效
标题翻译：酪氨酰-tRNA合成酶变体
公开(公告)号：US20050084856A1
公开(公告)日：2005-04-21
申请号：US10483636
申请日：2002-01-11
申请人： Daisuke Kiga , Kensaku Sakamoto , Ichiro Hirao , Shigeyuki Yokoyama
发明人： Daisuke Kiga , Kensaku Sakamoto , Ichiro Hirao , Shigeyuki Yokoyama
IPC分类号： C12N1/21 , C12N9/00 , C12Q1/68 , C07H21/04 , C12N9/22 , C12N15/74
CPC分类号： C12N9/93 , G01N2333/9125
摘要： Tyrosyl-tRNA synthetases having a modified amino acid sequence whereby unnatural amino acids can be more efficiently incorporated than original natural amino acids. More specifically, tyrosyl-tRNA synthetases having been modified at the amino acid(s) at the 37- and/or 195-positions. A method of modifying tyrosyl-tRNA synthetase characterized in that the position of an amino acid to be modified is determined based on the 3-D structure of a tyrosyl-AMP and of tyrosyl synthetase complex and an amino acid to which this enzyme binds.
摘要翻译：具有修饰氨基酸序列的酪氨酰-tRNA合成酶，由此可以比原始天然氨基酸更有效地掺入非天然氨基酸。更具体地，已经在37位和/或195位的氨基酸修饰的酪氨酰-tRNA合成酶。一种修饰酪氨酰-tRNA合成酶的方法，其特征在于基于酪氨酰-PAMP和酪氨酰合成酶复合物和该酶结合的氨基酸的3-D结构确定待修饰的氨基酸的位置。

4. 发明授权

US07393670B2 Tyrosyl-tRNA synthetase variants 失效
标题翻译：酪氨酰-tRNA合成酶变体
公开(公告)号：US07393670B2
公开(公告)日：2008-07-01
申请号：US10483636
申请日：2002-01-11
申请人： Daisuke Kiga , Kensaku Sakamoto , Ichiro Hirao , Shigeyuki Yokoyama
发明人： Daisuke Kiga , Kensaku Sakamoto , Ichiro Hirao , Shigeyuki Yokoyama
IPC分类号： C12N9/00 , C07K1/00
CPC分类号： C12N9/93 , G01N2333/9125
摘要： Tyrosyl-tRNA synthetases having a modified amino acid sequence whereby unnatural amino acids can be more efficiently incorporated than original natural amino acids. More specifically, tyrosyl-tRNA synthetases having been modified at the amino acid(s) at the 37- and/or 195-positions. A method of modifying tyrosyl-tRNA synthetase characterized in that the position of an amino acid to be modified is determined based on the 3-D structure of a tyrosyl-AMP and of tyrosyl synthetase complex and an amino acid to which this enzyme binds.
摘要翻译：具有修饰氨基酸序列的酪氨酰-tRNA合成酶，由此可以比原始天然氨基酸更有效地掺入非天然氨基酸。更具体地，已经在37位和/或195位的氨基酸修饰的酪氨酰-tRNA合成酶。一种修饰酪氨酰-tRNA合成酶的方法，其特征在于基于酪氨酰-PAMP和酪氨酰合成酶复合物和该酶结合的氨基酸的3-D结构确定待修饰的氨基酸的位置。

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