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    Sucrose phosphate synthetase isolated from maize
    2.
    发明授权
    Sucrose phosphate synthetase isolated from maize 失效
    从玉米分离的蔗糖磷酸合成酶

    公开(公告)号：US5714365A

    公开(公告)日：1998-02-03

    申请号：US376764

    申请日：1995-01-23

    申请人： Charles Van Assche ,  Danielle Lando ,  Jean Michel Bruneau ,  Toni Alois Voelker ,  Monica Gervais

    发明人： Charles Van Assche ,  Danielle Lando ,  Jean Michel Bruneau ,  Toni Alois Voelker ,  Monica Gervais

    IPC分类号： C07K16/40 ,  C12N9/10 ,  C12N15/82 ,  C12N9/12 ,  C12N15/00 ,  C12P19/12

    CPC分类号： C12N9/1029 ,  C07K16/40 ,  C12N15/8245 ,  C12N15/8261 ,  C12N15/8271 ,  C12N9/1066

    摘要： A protein having sucrose phosphate synthetase (SPS) activity is isolated from plants, preferably maize. The protein has a molecular weight of 110-130 dK and contains at least one peptide selected from Thv Trp Ile Lys, Try Val Val Glu Leu Ala Arg, Ser Met Pro Pro Ile Trp Ala Glu Val Met Arg, Leu Arg Pro Asp Gln Asp Try Leu Met His Ile Ser His Arg and Trp Ser His Asp Gly Ala Arg. Isolation is carried out by obtaining an extract from the plant by grinding, centrifugation and filtration; enriching the extract in SPS protein by precipitation in an appropriate solvent such as polyethylene glycol, centrifugation and solubilization of the precipitate obtained in a buffer solution; subjecting the protein thus obtained to low pressure anion exchange chromatography, chromatography on heparin Sepharose and high pressure anion exchange chromatography; and purifying the active fractions obtained by passage through two high pressure chromatography columns. Hybridomas and monoclonal antibodies are prepared from an antigen resulting from high pressure anion exchange chromatography above, antibodies directed specifically against SPS are selected and the antibodies are used to purify the SPS obtained previously. Complementary DNA coding for the SPS is prepared and used to modify expression of the SPS in plant cells.

    摘要翻译： 从植物，优选玉米中分离具有蔗糖磷酸合成酶（SPS）活性的蛋白质。 该蛋白质的分子量为110-130dK，并含有至少一种选自Thv Trp Ile Lys，Try Val Val Glu Leu Ala Arg，Ser Met Pro Pro Ile Trp Ala Glu Val Met Arg，Leu Arg Pro Asp Gln Asp 尝试Leu Met他的Ile Ser他的Arg和Trp Ser他的Asp Gly Ala Arg。 通过研磨，离心和过滤获得植物提取物进行分离; 通过在合适的溶剂如聚乙二醇中沉淀，使SPS蛋白中的提取物富集，离心并溶解在缓冲溶液中得到的沉淀; 对所得蛋白质进行低压阴离子交换色谱，肝素琼脂糖凝胶色谱和高压阴离子交换层析; 并纯化通过两个高压色谱柱获得的活性级分。 杂交瘤和单克隆抗体由上述高压阴离子交换层析得到的抗原制备，选择特异性针对SPS的抗体，并使用抗体纯化以前获得的SPS。 制备SPS编码的互补DNA，并用于修饰植物细胞中SPS的表达。

    Modification of sucrose phosphate synthase in plants
    6.
    发明授权
    Modification of sucrose phosphate synthase in plants 失效
    植物中蔗糖磷酸合成酶的修饰

    公开(公告)号：US5981852A

    公开(公告)日：1999-11-09

    申请号：US718777

    申请日：1996-09-24

    申请人： Charles Van Assche ,  Danielle Lando ,  Jean Michel Bruneau ,  Toni Alois Voelker ,  Monica Gervais

    发明人： Charles Van Assche ,  Danielle Lando ,  Jean Michel Bruneau ,  Toni Alois Voelker ,  Monica Gervais

    IPC分类号： C07K16/40 ,  C12N9/10 ,  C12N15/82 ,  A01H5/00 ,  C12N5/14

    CPC分类号： C07K16/40 ,  C12N15/8245 ,  C12N15/8261 ,  C12N15/8271 ,  C12N9/1029 ,  C12N9/1066

    摘要： Sucrose phosphate synthase (SPS), a process for its preparation, its DNA, and utilization of DNA encoding SPS to modify the expression of SPS in the plant cells are provided. Methods and compositions for producing SPS protein and DNA encoding SPS from a number of sources are provided. Also provided are transgenes comprising DNA encoding SPS and methods for the production of transgenic plant cells and plants which have extra SPS activity, and thus an increased capacity to synthesize sucrose. Transgenic plant cells and plants with the additional SPS activity exhibit altered carbon partitioning, growth and development, and yield under a variety of conditions.

    摘要翻译： 提供了蔗糖磷酸合成酶（SPS），其制备方法，其DNA，以及编码SPS的DNA修饰植物细胞中SPS表达的应用。 提供了从许多来源生产SPS蛋白和编码SPS的DNA的方法和组合物。 还提供了包含编码SPS的DNA的转基因和用于产生具有额外的SPS活性的转基因植物细胞和植物的方法，因此增加了合成蔗糖的能力。 具有附加SPS活性的转基因植物细胞和植物在各种条件下表现出改变的碳分配，生长和发育以及产量。