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    Nucleic Acid Assembly System
    2.
    发明申请
    Nucleic Acid Assembly System 审中-公开
    核酸装配系统

    公开(公告)号：US20140303036A1

    公开(公告)日：2014-10-09

    申请号：US14359358

    申请日：2012-11-23

    Applicant: DSM IP ASSETS B.V.

    Inventor： Johannes Andries Roubos ,  Bernard Meijrink ,  Richard Kerkman ,  Ben Den Dulk

    IPC: C12N15/10

    CPC classification number: C12N15/1082 ,  C12N15/1027 ,  C12N15/1093 ,  C12N15/52 ,  C12N15/80 ,  C12N15/905

    Abstract: The present invention relates to a method for the preparation of a library of host cells, a plurality of which comprise an assembled polynucleotide at a target locus, which method comprises: (a) providing a plurality of polynucleotides comprising two or more polynucleotide subgroups, wherein: (i) a plurality of polynucleotides in each polynucleotide subgroup comprises sequence encoding a peptide or polypeptide and/or a regulatory sequence; (ii) a plurality of peptides or polypeptides encoded by, or a plurality of regulatory sequences comprised within, each polynucleotide subgroup share an activity and/or function; (iii) at least one polynucleotide subgroup comprises at least two non-identical polynucleotide species; (iv) a plurality of polynucleotides of each polynucleotide subgroup comprises sequence enabling homologous recombination with a plurality of polynucleotides from one or more other polynucleotide subgroups; and (v) a plurality of polynucleotides in two polynucleotide subgroups comprise a nucleotide sequence enabling homologous recombination with a target locus in host cells; and (b) assembling the plurality of polynucleotides at the target locus by homologous recombination in vivo in host cells, thereby to generate a library of host cells, a plurality of which comprise an assembled polynucleotide at the target locus. The assembled polynucleotides may be recovered, thereby to prepare a library of nucleic acids.

    Abstract translation: 本发明涉及一种用于制备宿主细胞文库的方法，其多个在靶位点包含组装的多核苷酸，该方法包括：（a）提供多个包含两个或更多个多核苷酸亚基的多核苷酸，其中 （i）每个多核苷酸亚组中的多个多核苷酸包含编码肽或多肽和/或调节序列的序列; （ii）每个多核苷酸亚组中由多个或多个编码的多肽或多肽调节序列共享活性和/或功能; （iii）至少一个多核苷酸亚组包含至少两个不相同的多核苷酸种类; （iv）每个多核苷酸亚组的多个多核苷酸包括能够与来自一个或多个其它多核苷酸亚组的多个多核苷酸进行同源重组的序列; 和（v）两个多核苷酸亚组中的多个多核苷酸包含能够与宿主细胞中的靶基因座进行同源重组的核苷酸序列; 和（b）通过在宿主细胞中的体内同源重组在靶位点组装多个多核苷酸，从而产生宿主细胞文库，其多个包含靶标位点处的组装的多核苷酸。 可以回收组装的多核苷酸，从而制备核酸文库。

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