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    TRANSFORMATION METHOD OF SUGAR BEET PROTOPLASTS BY TALEN PLATFORM TECHNOLOGY
    2.
    发明公开
    TRANSFORMATION METHOD OF SUGAR BEET PROTOPLASTS BY TALEN PLATFORM TECHNOLOGY 审中-公开
    台糖平台技术转化甜菜原生质体的方法

    公开(公告)号：EP3155001A1

    公开(公告)日：2017-04-19

    申请号：EP15730110.2

    申请日：2015-06-12

    申请人： SESVanderHave N.V.

    发明人： WEYENS, Guy ,  LEFÈBVRE, Marc ,  THAMMANNAGOWDA, Shivegowda ,  LOMMEL, Murielle ,  SONG, Luo

    IPC分类号： C07K14/415 ,  C12N15/82

    CPC分类号： C12N15/8213 ,  C12N9/88

    摘要： A method for transformation of sugar beet protoplasts includes obtaining protoplasts from stomatal guard cells isolated from a sugar beet plant. The protoplasts are transformed with a nucleic acid construct including a nucleotide sequence of interest and Transcription Activator-Like Effector Nucleases (TALEN) or one or more vectors including sequences encoding these Transcription Activator-Like Effector Nucleases (TALEN)sequences. The TALEN target and process a target sequence and replace the target sequence through homologous recombination with the nucleic acid construct including the nucleotide sequence of interest, -possibly applying to an in vitroculture of the protoplasts, a medium that is toxic, preferably lethal to the in vitroculture of the protoplasts. Sugar beet plants are regenerated from the cell culture, preferably from the surviving protoplasts having integrated the nucleic acid construct including the sequence of interest that possibly renders the transformed cell resistant to the toxic activity of the applied medium.

    摘要翻译： 用于转化甜菜原生质体的方法包括以下步骤： - 从由甜菜植物分离的气孔保卫细胞获得原生质体， - 用包含目的核苷酸序列和转录激活物样效应物核酸酶的核酸构建体转化所述原生质体（ TALEN）或包含编码这些转录激活物样效应物核酸酶（TALEN）序列的序列的一种或多种载体，其中这些TALEN靶向并加工靶序列并通过与包含目标核苷酸序列的核酸构建体同源重组来替换该靶序列 ，可能适用于这些原生质体的培养物，有毒的，优选对原生质体的培养物致死的培养基，以及 - 从细胞培养物中再生甜菜植物，优选来自已整合有核酸构建体的存活原生质体 包括可能的兴趣序列 使得转化的细胞对所施用的培养基的毒性具有抗性。