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    • 4. 发明公开
    • VERFAHREN ZUR HERSTELLUNG EINER NUKLEINSÄURE- BIBLIOTHEK MIT MINDESTENS ZWEI BENACHBARTEN VARIABLEN CODON- TRIPLETTS
    • 制造方法的核酸库,至少两个相邻的可变密码子三联
    • EP2748319A1
    • 2014-07-02
    • EP13704605.8
    • 2013-02-14
    • Fuhrmann, Markus
    • Fuhrmann, Markus
    • C12N15/10C12N15/66
    • C12N15/1093C12N15/10C12N15/66C40B50/08
    • The invention relates to a method for producing a DNA sequence for a nucleic acid library having a plurality of elements. The synthesized DNA sequence comprises at least three regions. The first region and the third region in each element each have defined sequences (D1, D2) that are identical in each element of the nucleic acid library. In a middle second region, at least two adjacent variable codon triplets are arranged. The following method steps are carried out: First, oligonucleotides (O1-x) of a first oligonucleotide library (O1b) are hybridized with a first largely complementary oligonucleotide (O2) to form a first double-stranded DNA library (O1xO2). Furthermore, oligonucleotides (O3-x) of a second oligonucleotide library (O3b) are hybridized with a second largely complementary oligonucleotide (O4) to form a second double-stranded DNA library (O3xO4). Then the first double-stranded DNA library (O1xO2) and the second double-stranded DNA library (O3xO4) are ligated to form a double-stranded DNA ligation product (LPx). The double-stranded DNA ligation product comprises the following sub-regions: a defined first region (D1) of the nucleic acid library, at least one first variable codon triplet (Tv1), a spacer sequence (S), at least one second variable codon triplet (Tv2), and a defined second region (D2) of the nucleic acid library. After the spacer sequence (S) has been removed, a library of double-stranded DNA product (Px) comprising the following sub-regions is obtained: a defined first region (D1) of the nucleic acid library, at least one first variable codon triplet (Tv1), at least one second variable codon triplet (Tv2), and a defined second region (D2) of the nucleic acid library.
    • 7. 发明公开
    • PEPTIDE LIBRARY PRODUCTION METHOD, PEPTIDE LIBRARY, AND SCREENING METHOD
    • PEPTIDBIBLIOTHEK-HERSTELLUNGSVERFAHREN,PEPTIDBIBIIHEK UND SCREENING-VERFAHREN
    • EP2647720A1
    • 2013-10-09
    • EP11845623.5
    • 2011-12-05
    • The University of Tokyo
    • SUGA, HiroakiMORIMOTO, Jumpei
    • C12P21/00C12N15/09C40B40/06C40B40/10G01N33/15G01N33/50G01N33/573C12Q1/34
    • B01J19/0046B01J2219/00725C07K7/08C07K14/705C12N15/1062C12N15/67C12Q1/34C40B50/06C40B50/08
    • Object is to provide a method of constructing a library of peptides having, at a desired position in the random sequence of each peptide, an amino acid having a portion capable of binding to a target. The invention provides a method of producing a library of peptides having, at a designated position in the random sequence of each peptide, a special amino acid having a portion capable of binding to a target, including (i) preparing a library of mRNAs having, in the mRNA sequence coding for a random amino acid sequence, a base sequence having an altered codon encoding the special amino acid, (ii) preparing an aminoacyl tRNA with the special amino acid linked to a tRNA encoded by the altered codon, and (iii) translating the mRNAs by using a cell-free translation system containing the aminoacyl tRNA to obtain a library of peptides having, in the random sequence thereof, a predetermined special amino acid.
    • 目的是提供构建肽的文库的方法,其在每个肽的随机序列中的期望位置具有具有能够结合靶标的部分的氨基酸。 本发明提供一种生产肽文库的方法,其在每个肽的随机序列中的指定位置具有具有能与靶标结合的部分的特殊氨基酸,包括(i)制备mRNA文库, 在编码随机氨基酸序列的mRNA序列中,具有编码特殊氨基酸的改变的密码子的碱基序列,(ii)用与被改变的密码子编码的tRNA连接的特殊氨基酸制备氨酰基tRNA,和(iii )通过使用含有氨酰基tRNA的无细胞翻译系统来翻译mRNA,以获得在其随机序列中具有预定的特殊氨基酸的肽文库。