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    Plant cell comprising mutation introduced in target DNA, and method for producing the plant cell
    3.
    发明授权
    Plant cell comprising mutation introduced in target DNA, and method for producing the plant cell 有权

    公开(公告)号：US10301636B2

    公开(公告)日：2019-05-28

    申请号：US15121458

    申请日：2015-02-24

    申请人： NATIONAL RESEARCH AND DEVELOPMENT AGENCY NATIONAL AGRICULTURE AND FOOD RESEARCH ORGANIZATION

    发明人： Masaki Iwakami ,  Seiichi Toki

    IPC分类号： C12N15/82

    摘要： It has been found that a marker gene, which is inserted in the genomic DNA via homologous recombination, and to both ends of which nuclease recognition sites are added, can be removed from a plant cell by using a corresponding nuclease, and further that the nuclease recognition sites can also be removed without leaving any trace by matching sequences of at least 30 nucleotides adjacent to the recognition sites. Moreover, in a method for introducing a mutation into a target DNA on the genome of a plant cell via homologous recombination, it is made possible to: stably select a plant cell, in which the mutation is introduced, based on an expression of a marker gene; further, to remove an unnecessary sequence such as the marker gene from the selected cell; and to introduce only a required mutation into the target DNA.

    PLANT CELL COMPRISING MUTATION INTRODUCED IN TARGET DNA, AND METHOD FOR PRODUCING THE PLANT CELL
    4.
    发明申请
    PLANT CELL COMPRISING MUTATION INTRODUCED IN TARGET DNA, AND METHOD FOR PRODUCING THE PLANT CELL 审中-公开
    包含目标DNA引入突变的植物细胞，以及生产植物细胞的方法

    公开(公告)号：US20170058285A1

    公开(公告)日：2017-03-02

    申请号：US15119425

    申请日：2015-02-19

    申请人： NATIONAL RESEARCH AND DEVELOPMENT AGENCY NATIONAL AGRICULTURE AND FOOD RESEARCH ORGANIZATION

    发明人： Ayako YOKOI ,  Seiichi TOKI

    IPC分类号： C12N15/82

    CPC分类号： C12N15/8213 ,  C12N15/8209 ,  C12N15/8216

    摘要： A method for producing a plant cell comprising a mutation introduced in a target DNA comprises: a step of introducing into plant cells a DNA construct comprising a DNA homologous to a target DNA, wherein a desired mutation is introduced and a piggyBac transposon containing a marker gene is inserted in the homologous DNA; a step of selecting a plant cell, in which the mutation and the piggyBac transposon are introduced in the target DNA via homologous recombination, based on an expression of the marker gene; and a step of removing the piggyBac transposon from the target DNA by constitutively expressing a piggyBac transposase in the cell selected in the above step.

    摘要翻译： 用于生产向靶DNA引入突变的植物细胞的方法包括：向植物细胞中引入含有与靶DNA同源的DNA的DNA构建体，其中含有标记基因的PiggyBac转座子已经被 引入同源DNA并引入了所需的突变; 通过突变的同源重组和PiggyBac转座子，以标记基因的表达作为指标选择已经引入了靶DNA的植物细胞的步骤; 以及通过在上述步骤中选择的细胞中构建表达PiggyBac转座子从目标DNA中消除PiggyBac转座子的步骤。