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    FRT recombination sites and methods of use
    73.
    发明授权
    FRT recombination sites and methods of use 有权
    FRT重组位点和使用方法

    公开(公告)号：US07736897B2

    公开(公告)日：2010-06-15

    申请号：US11487300

    申请日：2006-07-14

    申请人： Yumin Tao ,  Dennis L. Bidney ,  William J. Gordon-Kamm ,  Leszek A. Lyznik

    发明人： Yumin Tao ,  Dennis L. Bidney ,  William J. Gordon-Kamm ,  Leszek A. Lyznik

    IPC分类号： C12N15/87 ,  C12N15/82

    CPC分类号： C12N15/8213 ,  C12N15/102 ,  C12N15/1051 ,  C12N15/1093 ,  C40B40/08

    摘要： Methods and compositions using populations of randomized modified FRT recombination sites to identify, isolate and/or characterize modified FRT recombination sites are provided. The recombinogenic modified FRT recombination sites can be employed in a variety of methods for targeted recombination of polynucleotides of interest, including methods to recombine polynucleotides, assess promoter activity, directly select transformed organisms, minimize or eliminate expression resulting from random integration into the genome of an organism, such as a plant, remove polynucleotides of interest, combine multiple transfer cassettes, invert or excise a polynucleotide, and identify and/or characterize transcriptional regulating regions are also provided.

    摘要翻译： 提供了使用随机修饰的FRT重组位点群的方法和组合来鉴定，分离和/或表征修饰的FRT重组位点。 重组生成修饰的FRT重组位点可用于多种感兴趣多核苷酸靶向重组方法，包括重组多核苷酸的方法，评估启动子活性，直接选择转化的生物体，最小化或消除由随机整合到基因组中的表达 生物体如植物，去除感兴趣的多核苷酸，组合多个转移盒，反转或切割多核苷酸，以及鉴定和/或表征转录调节区。

    Mobilization of viral genomes from T-DNA using site-specific recombination systems
    76.
    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：US07364902B2

    公开(公告)日：2008-04-29

    申请号：US11400293

    申请日：2006-04-07

    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： C12N15/82 ,  C12N5/04 ,  C12N5/10 ,  A01H1/00 ,  C07H21/00 ,  C07H21/02

    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.

    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。

    Method for the integration of foreign DNA into eukaryotic genomes
    79.
    发明授权
    Method for the integration of foreign DNA into eukaryotic genomes 有权
    将外源DNA整合到真核生物基因组中的方法

    公开(公告)号：US07223601B2

    公开(公告)日：2007-05-29

    申请号：US10353445

    申请日：2003-01-29

    申请人： Christopher L. Baszczynski ,  Leszek Alexander Lyznik ,  William J. Gordon-Kamm ,  Xueni Guan ,  Aragula Gururaj Rao ,  Laura A. Tagliani

    发明人： Christopher L. Baszczynski ,  Leszek Alexander Lyznik ,  William J. Gordon-Kamm ,  Xueni Guan ,  Aragula Gururaj Rao ,  Laura A. Tagliani

    IPC分类号： C12N15/87

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

    摘要： Compositions and methods for introducing a DNA of interest into a genomic target site are provided. In particular, the methods and compositions involve the use of a combination of target sites for two site specific recombinases and expression of a chimeric recombinase with dual target site specificity. Thus, the compositions comprise novel site-specific recombinases with specificities to multiple target sites, and nucleotide sequences and expression cassettes encoding these recombinases or target sites. The methods involve transforming a eukaryotic cell having target sites for the novel recombinase with a DNA of interest that is flanked by corresponding target sites. Expression of the recombinase results in integration of the DNA of interest into the genome of the cell. The compositions and methods of the invention have use in the construction of stably transformed eukaryotic cells, and in particular, plant cells. The methods result in the efficient targeted genomic integration of DNA by site-specific recombination.

    摘要翻译： 提供了将目的DNA引入基因组靶位点的组合物和方法。 特别地，所述方法和组合物涉及使用两个位点特异性重组酶的靶位点的组合和具有双重靶位点特异性的嵌合重组酶的表达。 因此，组合物包含对多个靶位点具有特异性的新的位点特异性重组酶，以及编码这些重组酶或靶位点的核苷酸序列和表达盒。 所述方法包括用具有相应靶位点侧翼的感兴趣的DNA转化具有新重组酶靶位点的真核细胞。 重组酶的表达导致感兴趣的DNA整合到细胞的基因组中。 本发明的组合物和方法用于构建稳定转化的真核细胞，特别是植物细胞。 该方法通过位点特异性重组导致DNA的有效靶向基因组整合。

    Mobilization of viral genomes from T-DNA using site-specific recombination systems
    80.
    发明授权
    Mobilization of viral genomes from T-DNA using site-specific recombination systems 有权
    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：US07179599B2

    公开(公告)日：2007-02-20

    申请号：US10444664

    申请日：2003-05-23

    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： C12Q1/68 ,  C12N1/12 ,  C12N1/20 ,  C12N15/00 ,  C12N15/09

    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 。