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公开(公告)号：US08715984B2
公开(公告)日：2014-05-06
申请号：US12866720
申请日：2009-02-03
CPC分类号： C12N15/67 , C12N15/11 , C12N2310/33 , C12P21/02 , C12Q1/25 , C12Q1/48 , G01N2500/00
摘要： In the method for introducing a noncanonical amino acid residue into a desired position in a protein, the structure of tRNA is so modified as to have improved affinity for aminoacyl-tRNA synthetase or improved specificity to aminoacyl-tRNA synthetase. An unnatural base is contained at any position in tRNA, whereby the efficiency of aminoacylation of the tRNA with a noncanonical amino acid can be improved.
摘要翻译： 在将非氨基酸氨基酸残基引入蛋白质中所需位置的方法中，对tRNA的结构进行了修饰，以改善对氨基-tRNA合成酶的亲和力或提高了对氨基-tRNA合成酶的特异性。 在tRNA中的任何位置都含有非天然碱基，从而可以提高tRNA与非科学氨基酸的氨酰化效率。
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2.发明授权Mutant SepRS, and method for site-specific introduction of phosphoserine into protein using the same 失效突变体SepRS，以及使用其将磷酸丝氨酸位点特异性引入蛋白质的方法公开(公告)号：US08178301B2
公开(公告)日：2012-05-15
申请号：US12318344
申请日：2008-12-24
摘要： A mutant SepRS which is suitable for a site-specific introduction of phosphoserine into a protein is prepared by analyzing the structure and functions of a phosphoseryl-tRNA synthetase (SepRS) derived from an archaebacterium. A mutant SepRS composed of an amino acid sequence depicted in SEQ ID NO:2, in which any one or more of glutamic acids at position-418 and position-420 and threonine at position-423 are substituted with other amino acid, and having enhanced binding affinity with a suppressor tRNA as compared with a wild type phosphoseryl-tRNA synthetase (SepRS) composed of an amino acid sequence depicted in SEQ ID NO:2 is provided.
摘要翻译： 通过分析源自古细菌的磷酰基-tRNA合成酶（SepRS）的结构和功能，制备适合于将磷酸丝氨酸位点特异性引入蛋白质的突变体SepRS。 由SEQ ID NO：2所示的氨基酸序列构成的突变体SepRS，其中418位和420位的谷氨酸和423位的苏氨酸中的任何一种或多种被其它氨基酸取代，并具有增强的 提供了与由SEQ ID NO：2所示的氨基酸序列组成的野生型磷酰基-tRNA合成酶（SepRS）相比，与抑制子tRNA的结合亲和性。
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3.发明申请Mutant SepRS, and method for site-specific introduction of phosphoserine into protein using the same 失效突变体SepRS，以及使用其将磷酸丝氨酸位点特异性引入蛋白质的方法公开(公告)号：US20090233290A1
公开(公告)日：2009-09-17
申请号：US12318344
申请日：2008-12-24
摘要： A mutant SepRS which is suitable for a site-specific introduction of phosphoserine into a protein is prepared by analyzing the structure and functions of a phosphoseryl-tRNA synthetase (SepRS) derived from an archaebacterium. A mutant SepRS composed of an amino acid sequence depicted in SEQ ID NO:2, in which any one or more of glutamic acids at position-418 and position-420 and threonine at position-423 are substituted with other amino acid, and having enhanced binding affinity with a suppressor tRNA as compared with a wild type phosphoseryl-tRNA synthetase (SepRS) composed of an amino acid sequence depicted in SEQ ID NO:2 is provided.
摘要翻译： 通过分析源自古细菌的磷酰基-tRNA合成酶（SepRS）的结构和功能，制备适合于将磷酸丝氨酸位点特异性引入蛋白质的突变体SepRS。 由SEQ ID NO：2所示的氨基酸序列构成的突变型SepRS，其中418位和420位的谷氨酸和423位的苏氨酸中的任何一种或多种被其它氨基酸取代，并具有增强的 提供了与由SEQ ID NO：2所示的氨基酸序列组成的野生型磷酰基-tRNA合成酶（SepRS）相比，与抑制子tRNA的结合亲和性。
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4.发明授权Nucleic acid base analogs with quenching and fluorescent activities and applications thereof 有权具有淬灭和荧光活性的核酸碱基类似物及其应用公开(公告)号：US09285319B2
公开(公告)日：2016-03-15
申请号：US13642111
申请日：2011-04-21
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04 , G01N21/64 , C07H19/044
CPC分类号： G01N21/6486 , C07H19/044 , C12Q1/6818 , C12Q2525/117
摘要： It is an object of the present invention to provide quenching or fluorescent nucleic acid base analogs and applications thereof. The quencher of the present invention has a 2-nitropyrrole structure represented by Formula I:[Formula 1] (in Formula I, R1 and R2 are groups independently selected from the group consisting of: ribose and deoxyribose; hydrogen, hydroxyl and SH groups, and halogens; substituted or unsubstituted alkyl, alkenyl, and alkynyl groups each having 2 to 10 carbon atoms; one or more five-membered heterocyclic rings, one or more six-membered heterocyclic rings, and one or more fused heterocyclic rings, these heterocylic rings containing nitrogen or sulfur, and one or more aromatic rings; sugars, sugar chains, amino acids, and peptides; and fluorescent molecules linked via linkers).
摘要翻译： 本发明的目的是提供淬灭或荧光核酸碱基类似物及其应用。 本发明的猝灭剂具有由式I表示的2-硝基吡咯结构：[式1]（式I中，R 1和R 2分别独立地选自：核糖和脱氧核糖;氢，羟基和SH基团， 卤素;取代或未取代的各自具有2-10个碳原子的烷基，烯基和炔基;一个或多个五元杂环，一个或多个六元杂环和一个或多个稠合杂环，这些杂环 含有氮或硫，以及一个或多个芳环;糖，糖链，氨基酸和肽;以及通过接头连接的荧光分子）。
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公开(公告)号：US08686130B2
公开(公告)日：2014-04-01
申请号：US13500303
申请日：2010-10-06
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui
CPC分类号： C07D471/04 , C07D473/32 , C07H19/173 , C07H19/23 , C09K11/06 , C09K2211/1074
摘要： The present invention relates to novel unnatural fluorescent nucleic acid bases, that is, a purine base, a 1-deazapurine base, and a 1,7-deazapurine base each having a functional group which consists of two or more heterocyclic moieties linked together, at the 6-position thereof (the 6-position of purine ring). The present invention also relates to a compound containing the unnatural base, a derivative thereof, and a nucleic acid containing a nucleotide having the unnatural base. The present invention also relates to a method of preparing the nucleic acid. The unnatural base of the present invention has excellent fluorescence characteristics and also has excellent properties as a universal base.
摘要翻译： 本发明涉及新型非天然荧光核酸碱基，即嘌呤碱，1-脱氮嘌呤碱和1,7-deazapurine碱，其各自具有由两个或多个连接在一起的杂环基部分组成的官能团，在 其6-位（嘌呤环的6-位）。 本发明还涉及含有非天然碱，其衍生物和含有具有非天然碱基的核苷酸的核酸的化合物。 本发明还涉及制备核酸的方法。 本发明的非天然碱具有优异的荧光特性，也具有优异的作为通用基材的特性。
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6.发明授权公开(公告)号：US08642291B2
公开(公告)日：2014-02-04
申请号：US13470752
申请日：2012-05-14
CPC分类号： C12P21/02 , C12N9/93 , C12N2510/02
摘要： Producing proteins incorporating non-natural amino acids can involve introducing genes into and knocking inherent genes out of eukaryote-type cells. Genes to be introduced include genes encoding eukaryote-type aminoacyl tRNA synthetase mutants having enhanced specificity to non-natural amino acids, compared with specificity to similar natural amino acids, and tRNA genes for non-natural amino acids capable of binding to the non-natural amino acids in the presence of the eukaryote-type aminoacyl tRNA synthetase mutants. Inherent genes to be knocked out include genes encoding aminoacyl tRNA synthetase having specificity to natural amino acids and tRNA genes capable of binding to the natural amino acids in the presence of the inherent aminoacyl tRNA synthetase.
摘要翻译： 产生含有非天然氨基酸的蛋白质可以涉及将基因导入真核细胞型细胞内并敲入固有基因。 要引入的基因包括具有对非天然氨基酸的特异性增强的真核生物型氨基酰基tRNA合成酶突变体的基因，与对相似天然氨基酸的特异性相比，以及能够结合非天然氨基酸的非天然氨基酸的tRNA基因 在真核生物型氨酰tRNA合成酶突变体存在下的氨基酸。 要敲除的固有基因包括编码具有对天然氨基酸具有特异性的氨基酰基tRNA合成酶的基因和能够在固有氨基tRNA合成酶存在下与天然氨基酸结合的tRNA基因。
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7.发明授权Extract of E. coli cells having mutation in ribosomal protein S12, and method for producing protein in cell-free system using the extract 失效在核糖体蛋白S12中具有突变的大肠杆菌细胞的提取物，以及使用提取物在无细胞系统中产生蛋白质的方法公开(公告)号：US08603774B2
公开(公告)日：2013-12-10
申请号：US11137395
申请日：2005-05-26
申请人： Namtip Chumpolkulwong , Chie Hori , Shigeyuki Yokoyama , Mikako Shirouzu , Takanori Kigawa , Kozo Ochi , Takeshi Hosaka
发明人： Namtip Chumpolkulwong , Chie Hori , Shigeyuki Yokoyama , Mikako Shirouzu , Takanori Kigawa , Kozo Ochi , Takeshi Hosaka
IPC分类号： C12P21/02
CPC分类号： C12P21/02
摘要： It is intended to provide an extract, kit and a process for synthesizing a protein in a cell-free system. The extract is characterized by being prepared from mutant Escherichia coli cells having a mutation in the ribosomal protein S12 gene. In a preferred embodiment, the mutation is such a mutation as conferring a resistance or dependence to streptomycin on E. coli whereby the reading efficiency of mRNA codon on ribosomes can be improved and thus the protein productivity can be elevated.
摘要翻译： 旨在提供在无细胞系统中合成蛋白质的提取物，试剂盒和方法。 提取物的特征在于由在核糖体蛋白S12基因中具有突变的突变型大肠杆菌细胞制备。 在一个优选的实施方案中，突变是这样的突变，其赋予对大肠杆菌对链霉素的抗性或依赖性，从而可以提高核糖体上的mRNA密码子的阅读效率，从而提高蛋白质生产力。
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8.发明授权Cell-free system for synthesis of proteins derived from cultured mammalian cells 有权用于合成来自培养的哺乳动物细胞的蛋白质的无细胞系统公开(公告)号：US08445232B2
公开(公告)日：2013-05-21
申请号：US13211280
申请日：2011-08-16
IPC分类号： C12P21/00
CPC分类号： C12P21/02 , C07K14/4702
摘要： Prepared is an extract composition having an improved protein synthetic activity in a cell-free protein synthesis system using a mammalian cultured cell extract. An eukaryotic translation initiation factor and/or translational regulator are added to a cell-free protein synthesis system comprising an extract prepared from cultured mammalian cells and a template mRNA. These factors are one or more selected from the group consisting of eukaryotic translation initiation factors 4E (eIF4E), 2 (eIF2) and 2B (eIF2B), and eukaryotic translational regulator p97.
摘要翻译： 制备的是在使用哺乳动物培养细胞提取物的无细胞蛋白质合成系统中具有改进的蛋白质合成活性的提取物组合物。 将真核翻译起始因子和/或翻译调节子添加到包含由培养的哺乳动物细胞和模板mRNA制备的提取物的无细胞蛋白质合成系统中。 这些因子是选自真核翻译起始因子4E（eIF4E），2（eIF2）和2B（eIF2B）以及真核翻译调节子p97中的一种或多种。
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9.发明申请公开(公告)号：US20120028330A1
公开(公告)日：2012-02-02
申请号：US13180209
申请日：2011-07-11
CPC分类号： C12N9/93
摘要： A polypeptide according to the present invention includes: an altered polypeptide obtained by altering an ArgRS, a CysRS, a MetRS, a GlnRS, a GluRS, a LysRS, a TyrRS, or a TrpRS so that an unnatural amino acid is recognized; and an editing polypeptide derived from a PheRS, a LeuRS, an IleRS, a ValRS, an AlaRS, a ProRS, or a ThrRS, the editing polypeptide having been either inserted between a Rossman-fold N domain and a Rossman-fold C domain that exist in the altered polypeptide, or bound to an N terminal of the altered polypeptide. Thus provided are a new aaRS that exhibits high substrate specificity to an unnatural amino acid and a technique that involves the use of such an aaRS.
摘要翻译： 根据本发明的多肽包括：通过改变ArgRS，CysRS，MetRS，GlnRS，GluRS，LysRS，TyrRS或TrpRS获得的改变的多肽，使得识别出非天然氨基酸; 和衍生自PheRS，LeuRS，IleRS，ValRS，AlaRS，ProRS或ThrRS的编辑多肽，所述编辑多肽已插入Rossman折叠N结构域和Rossman折叠C结构域之间， 存在于改变的多肽中，或者与改变的多肽的N末端结合。 因此提供了对非天然氨基酸表现出高底物特异性的新aaRS和涉及使用这种aaRS的技术。
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公开(公告)号：US07960543B2
公开(公告)日：2011-06-14
申请号：US11667146
申请日：2005-11-08
申请人： Ichiro Hirao , Shigeyuki Yokoyama
发明人： Ichiro Hirao , Shigeyuki Yokoyama
CPC分类号： C07H21/02
摘要： The object of the present invention is to provide a nucleoside or nucleotide having a 5-substituted-2-oxo(1H)pyridin-3-yl group as a base, as well as a method using the same.In one embodiment, the nucleoside or nucleotide of the present invention has a fluorescent dye selected from the group consisting of 5-FAM, 6-FAM, 5-TAMRA, 6-TAMRA, DANSYL, 5-HEX, 6-HEX, 5-TET, 6-TET, 5-ROX and 6-ROX or a quencher dye selected from the group consisting of DABCYL, BHQ1 and BHQ2, which is attached either directly or through a linker to the 5-position of the above base.
摘要翻译： 本发明的目的是提供具有5-取代-2-氧代（1H）吡啶-3-基作为碱的核苷或核苷酸，以及使用其的方法。 在一个实施方案中，本发明的核苷或核苷酸具有选自5-FAM，6-FAM，5-TAMRA，6-TAMRA，DANSYL，5-HEX，6-HEX，5- TET，6-TET，5-ROX和6-ROX或选自DABCYL，BHQ1和BHQ2的猝灭剂染料，其直接或通过接头连接到上述碱基的5位上。
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