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    Process for generating a variant library of DNA sequences
    3.
    发明授权
    Process for generating a variant library of DNA sequences 有权

    公开(公告)号：US09856470B2

    公开(公告)日：2018-01-02

    申请号：US12960061

    申请日：2010-12-03

    申请人： Thomas Greiner-Stoeffele ,  Claudia Feller ,  Marc Struhalla

    发明人： Thomas Greiner-Stoeffele ,  Claudia Feller ,  Marc Struhalla

    IPC分类号： C12Q1/68 ,  C12N15/10

    CPC分类号： C12N15/1093 ,  C12N15/102

    摘要： A method for generating a variant library of DNA sequences starting from at least one DNA starting sequence and including the steps: a) selecting at least two mutation sites in the starting sequence; b) dividing the DNA starting sequence into at least two sequence segments such that at least two of these sequence segments each contain at least one of the mutation sites; c) amplifying the sequence segments by polymerase chain reaction using at least five different oligonucleotides, where at (i) least one of the oligonucleotides can attach to each mutation site; (ii) at least two of the oligonucleotides can attach to at least one mutation site, and (iii) mutations are introduced, via mismatch positions, into the PCR amplificates by the oligonucleotides at the mutation sites where at least two mutations are introduced at at least one of the mutation sites; and d) linking the amplificates to give DNA sequences.

    Method From the Selection of Biomolecules From Biomolecules Variant Libraries
    6.
    发明申请
    Method From the Selection of Biomolecules From Biomolecules Variant Libraries 审中-公开
    从生物分子选择变异图书馆的方法

    公开(公告)号：US20080220518A1

    公开(公告)日：2008-09-11

    申请号：US10576684

    申请日：2004-10-22

    申请人： Thomas Greiner-Stoffele ,  Marc Struhalla

    发明人： Thomas Greiner-Stoffele ,  Marc Struhalla

    IPC分类号： C12N15/00 ,  C40B20/00 ,  C40B20/08

    CPC分类号： C12N15/1075

    摘要： The invention relates to a method from the selection of biomolecules from variant libraries, in particular of biocatalytically active biomolecules, comprising the steps: a) production of a variant library, b) division of the library into a number of compartments, which is smaller than the total number of variants in the variant library by a factor of at least 10, c) production and testing of the biomolecules in the individual compartments for a particular property, for example, a biocatalytic activity, d) selection of at least one compartment I in which there are biomolecules fulfilling the desired property, e) division of the partial library contained in the selected compartment into further compartments and f) n-fold repetition of the steps c) to e) until each compartment contains only one variant of the gene sequence coding for the biomolecule. In contrast to established methods which comprise mutagenesis and selection steps, said method starts with a large library in which the desired variant is contained from the outset.

    摘要翻译： 本发明涉及从变体文库，特别是生物催化活性生物分子选择生物分子的方法，其包括以下步骤：a）产生变体文库，b）将文库分离成若干个室，其小于 所述变体文库中的变体总数至少为10，c）生产和测试各个区室中特定性质的生物分子，例如生物催化活性，d）选择至少一个隔室I 其中存在满足所需性质的生物分子，e）将所选择的隔室中包含的部分文库划分成另外的隔室，以及f）步骤c）至e）的n倍重复，直到每个隔室仅含有该基因的一个变体 生物分子的序列编码。 与包括诱变和选择步骤的既定方法相反，所述方法从一开始就包含所需变体的大型文库开始。