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    • 4. 发明授权
    • Labeled protein and its producing method, labeling compound to be used in the method, and method for analyzing function of genes
    • 标记蛋白及其制备方法,方法中使用的标记化合物和分析基因功能的方法
    • US07041446B2
    • 2006-05-09
    • US09794128
    • 2001-02-28
    • Hiroshi YanagawaNaoto NemotoEtsuko Miyamoto
    • Hiroshi YanagawaNaoto NemotoEtsuko Miyamoto
    • C12Q1/68C07K1/13
    • C07H19/20C07H19/04C07K1/13
    • A protein having a labeling compound attached to its C-terminal, in which the compound comprises a label portion comprising a label substance and an acceptor portion comprising a compound having an ability of binding to a C-terminal of a synthesized protein when protein synthesis is carried out in a cell-free protein synthesis system or in a living cell is provided. Also, a method for producing the protein comprising the step of performing synthesis of a protein in a cell-free protein synthesis system or in a living cell in the presence of a labeling compound comprising a label portion comprising a label substance and an acceptor portion comprising a compound having an ability of binding to a C-terminal of a synthesized protein when protein synthesis is carried out in the cell-free protein synthesis system or in the living cell, the labeling compound being present at a concentration effective for the labeling compound to bind to the C-terminal of the synthesized protein is provided. Also, the labeling compound and a method for analyzing a function of a gene are further provided.
    • 一种具有附着于其C-末端的标记化合物的蛋白质,其中所述化合物包含标记物部分,所述标记物包含标记物质和受体部分,所述受体部分包含当蛋白质合成为蛋白质合成时具有与合成蛋白质C末端结合的能力的化合物 在无细胞蛋白质合成系统或活细胞中进行。 另外,制备蛋白质的方法包括以下步骤:在标记化合物存在下,在无细胞蛋白质合成系统或活细胞中进行蛋白质的合成,所述标记化合物包含标记物质和受体部分,所述标记物包含 当在无细胞蛋白质合成系统或活细胞中进行蛋白质合成时,具有与合成蛋白质C末端结合的能力的化合物,标记化合物以对标记化合物有效的浓度存在 提供了与合成蛋白质的C-末端结合。 此外,还提供了标记化合物和分析基因功能的方法。
    • 7. 发明申请
    • Cleavable assigned molecules and screening method using the same
    • 可拆分的分子和筛选方法使用相同
    • US20060210982A1
    • 2006-09-21
    • US10543682
    • 2004-02-02
    • Hiroshi YanagawaNobuhide DoiTetsuya NaganoHideaki Takashima
    • Hiroshi YanagawaNobuhide DoiTetsuya NaganoHideaki Takashima
    • C40B40/08
    • C40B30/04C07K14/00C12N15/1075
    • A method for screening a nucleic acid library for a nucleic acid encoding a protein that interacts with a target substance, which comprises: the step of producing a library of assigning molecules each comprising a protein and a nucleic acid encoding the protein linked to each other via a linker cleavable under a condition that does not change a nucleotide sequence of the nucleic acid; the step of mixing the library of assigning molecules and the target substance; the step of separating an assigning molecule binding to the target substance; the step of cleaving a linker of the separated assigning molecule under a condition that does not change a nucleotide sequence of the nucleic acid to release the nucleic acid; and the step of collecting the released nucleic acid. By this method, screening for an assigning molecule that specifically binds to a target substance can be conducted with high efficiency.
    • 一种用于筛选编码与靶物质相互作用的蛋白质的核酸的核酸文库的方法,该方法包括:产生分配分子的文库的步骤,每个分子包含蛋白质和编码相互连接的蛋白质的核酸通过 在不改变核酸的核苷酸序列的条件下可切割的接头; 分配分子的文库和目标物质的步骤; 分离与靶物质结合的分配分子的步骤; 在不改变核酸的核苷酸序列以释放核酸的条件下切割分离的分配分子的接头的步骤; 以及收集释放的核酸的步骤。 通过该方法,可以高效率地进行与目标物质特异性结合的分配分子的筛选。
    • 9. 发明授权
    • Molecule that homologizes genotype and phenotype and utilization thereof
    • 分子基因型和表型同源化及其利用
    • US06361943B1
    • 2002-03-26
    • US09284627
    • 1999-06-02
    • Hiroshi YanagawaNaoto NemotoEtsuko MiyamotoYuzuru Husimi
    • Hiroshi YanagawaNaoto NemotoEtsuko MiyamotoYuzuru Husimi
    • C12Q168
    • C12N15/11C12N15/1062
    • A molecule comprising a nucleic acid portion and a protein portion directly bound to said nucleic acid portion with a covalent bond, wherein said nucleic acid portion comprises a polymer of nucleoside, and said protein portion is encoded by said nucleic acid portion, and a method for constructing the molecule, which comprises (a) preparing a DNA containing a gene which has no termination codon, (b) transcribing the prepared DNA into RNA, (c) bonding a chimeric spacer composed of DNA and RNA to a 3′-terminal end of the obtained RNA, (d) bonding to a 3′-terminal end of the obtained bonded product, a nucleoside or a substance having a chemical structure analogous to that of a nucleoside, which can be covalently bound to an amino acid or substance having a chemical structure analogous to that of an amino acid, and (e) performing protein synthesis in a cell-free protein synthesis system using the obtained bonded product as mRNA to bond a nucleic acid portion containing the gene to a translation product of the gene. The molecule of the present invention is an extremely useful substance that can be used for evolutionary molecular engineering, i.e., modification of functional biopolymers such as enzymes, antibodies, and ribozymes, and creation of biopolymers having functions which cannot be found in living organisms.
    • 包含核酸部分和与所述核酸部分直接键合有共价键的蛋白质部分的分子,其中所述核酸部分包含核苷聚合物,所述蛋白质部分由所述核酸部分编码, 构建分子,其包括(a)制备含有不具有终止密码子的基因的DNA,(b)将制备的DNA转录成RNA,(c)将由DNA和RNA组成的嵌合间隔物结合至3'末端 的所得RNA,(d)与获得的结合产物的3'-末端结合,具有与核苷类似的化学结构的核苷或物质,其可共价结合于具有 与氨基酸类似的化学结构,和(e)使用获得的结合产物作为mRNA将含有该基因的核酸部分键合到无转化蛋白质合成系统中进行蛋白质合成 该基因的产物。 本发明的分子是可用于进化分子工程,即功能性生物聚合物如酶,抗体和核酶的修饰以及具有在活生物体中不能发现的功能的生物聚合物的非常有用的物质。