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    Mobilization of viral genomes from T-DNA using site-specific recombination systems
    81.
    发明授权
    Mobilization of viral genomes from T-DNA using site-specific recombination systems 失效
    使用位点特异性重组系统动员来自T-DNA的病毒基因组

    公开(公告)号：US06664108B1

    公开(公告)日：2003-12-16

    申请号：US09411826

    申请日：1999-10-01

    申请人： Christopher L. Baszczynski ,  Leszek Alexander Lyznik ,  William J. Gordon-Kamm ,  Xueni Guan

    发明人： Christopher L. Baszczynski ,  Leszek Alexander Lyznik ,  William J. Gordon-Kamm ,  Xueni Guan

    IPC分类号： C12N1582

    CPC分类号： C12N15/8213 ,  C07K2319/00 ,  C12N9/00 ,  C12N15/8203 ,  C12N15/8205 ,  C12N15/90

    摘要： The invention relates to methods and compositions for site-specific recombinase-mediated mobilization of viral replicons and associated DNAs of interest from T-DNA. The methods of the invention comprise Agrobacterium-mediated transfer of T-DNA to a plant cell, wherein the T-DNA contains a viral replicon flanked by directly repeated target sites for a site-specific recombinase and optionally a DNA of interest linked to the viral replicon. The DNA of interest may also contain a non-identical target site for the recombinase. An expression cassette for the site-specific recombinase is present on the T-DNA or the plant genome, or is transiently introduced into the plant cell. Expression of the site-specific recombinase in the plant cell results in excision of the viral replicon and the associated DNA of interest. The viral replicon and DNA of interest are then replicated to high copy number in the host plant cell. The compositions of the invention comprise nucleic acids, such as T-DNAs containing a viral DNA flanked by directly repeated target sites for a site-specific recombinase. The nucleic acids of the invention may additionally contain expression cassettes encoding the cognate site-specific recombinase for the target sites flanking the viral genome. The compositions of the invention further comprise Agrobacterium containing the nucleic acids of the invention. The compositions and methods of the invention have use in increasing the efficiency of agroinfection, providing high copy numbers of a DNA of interest for transient expression or for integration into a plant chromosome, and in simplifying the construction and stable maintenance of vectors for agroinfection and transformation.

    摘要翻译： 本发明涉及用于位点特异性重组酶介导的来自T-DNA的感兴趣的病毒复制子和相关DNA的动员的方法和组合物。 本发明的方法包括农杆菌介导的T-DNA向植物细胞的转移，其中T-DNA含有病毒复制子，其侧翼是用于位点特异性重组酶的直接重复的靶位点，以及任选的与病毒连接的感兴趣的DNA 复制品。 感兴趣的DNA还可以含有重组酶的不相同的靶位点。 用于位点特异性重组酶的表达盒存在于T-DNA或植物基因组上，或瞬时引入植物细胞。 位点特异性重组酶在植物细胞中的表达导致病毒复制子和感兴趣的相关DNA的切除。 然后将病毒复制子和感兴趣的DNA复制到宿主植物细胞中的高拷贝数。 本发明的组合物包含核酸，例如含有侧翼为用于位点特异性重组酶的直接重复靶位点的病毒DNA的T-DNA。 本发明的核酸可以另外含有编码针对病毒基因组侧翼的靶位点的同源位点特异性重组酶的表达盒。 本发明的组合物还包含含有本发明核酸的农杆菌。 本发明的组合物和方法用于增加农业感染的效率，为临时表达提供高拷贝数的目的DNA，或用于整合到植物染色体中，以及简化用于农业感染和转化的载体的构建和稳定维持 。