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    • 4. 发明申请
    • Down-regulation and silencing of allergen genes in transgenic peanut plants
    • 转基因花生植物中过敏原基因的下调和沉默
    • US20050114924A1
    • 2005-05-26
    • US10958324
    • 2004-10-06
    • Hortense DodoCharles ArntzenOlga ViquezKoffi Konan
    • Hortense DodoCharles ArntzenOlga ViquezKoffi Konan
    • C07K14/415C12N15/82A01H1/00
    • C12N15/8251C07K14/415C12N15/8242
    • An allergen-free transgenic peanut seed is produced by recombinant methods. Peanut plants are transformed with multiple copies of each of the allergen genes, or fragments thereof, to suppress gene expression and allergen protein production. Alternatively, peanut plants are transformed with peanut allergen antisense genes introduced into the peanut genome as antisense fragments, sense fragments, or combinations of both antisense and sense fragments. Peanut transgenes are under the control of the 35S promoter, or the promoter of the Ara h2 gene to produce antisense RNAs, sense RNAs, and double-stranded RNAs for suppressing allergen protein production in peanut plants. A full length genomic clone for allergen Ara h2 is isolated and sequenced. The ORF is 622 nucleotides long. The predicted encoded protein is 207 amino acids long and includes a putative transit peptide of 21 residues. One polyadenilation signal is identified at position 951. Six additional stop codons are observed. A promoter region was revealed containing a putative TATA box located at position−72. Homologous regions were identified between Ara h2, h6, and h7, and between Ara h3 and h4, and between Ara h1P41B and Ara h1P17. The homologous regions will be used for the screening of peanut genomic library to isolate all peanut allergen genes and for down-regulation and silencing of multiple peanut allergen genes.
    • 通过重组方法产生无过敏原的转基因花生种子。 每个变应原基因或其片段的多个拷贝转化花生植物以抑制基因表达和变应原蛋白质的产生。 或者,用作为反义片段,有义片段或反义片段和有义片段的组合引入花生基因组的花生过敏原反义基因转化花生植物。 花生转基因由35S启动子或Ara h2基因的启动子控制,以产生用于抑制花生植物中过敏原蛋白质产生的反义RNA,有义RNA和双链RNA。 分离和测序过敏原Ara h2的全长基因组克隆。 ORF为622个核苷酸长。 预测编码的蛋白质是207个氨基酸长,并且包含21个残基的推定转运肽。 在951位点识别出一种多聚增生信号。观察到另外六个终止密码子。 揭示了一个启动子区域,其中包含位于-72位置的假定TATA盒。 在Ara h2,h6和h7之间以及Ara h3和h4之间以及Ara h1P41B和Ara h1P17之间鉴定了同源区域。 同源区域将用于筛选花生基因组文库,以分离所有花生过敏原基因并降低多个花生过敏原基因的沉默。
    • 10. 发明授权
    • Gemini virus vectors for gene expression in plants
    • 双子座病毒载体,用于植物中的基因表达
    • US06392121B1
    • 2002-05-21
    • US09414276
    • 1999-10-07
    • Hugh S. MasonKenneth E. PalmerKathleen L. HefferonTsafrir S. MorCharles Arntzen
    • Hugh S. MasonKenneth E. PalmerKathleen L. HefferonTsafrir S. MorCharles Arntzen
    • C12N504
    • C12N15/8216C12N15/8203Y10S977/804
    • A gene amplification system based on plant viral genetic elements dramatically increases foreign protein production in plants. A safer and more economical production system for vaccines and antibodies in recombinant plants grown using agricultural practice is described. The high-level expression system uses the replicative process of a plant mastrevirus, exemplified by bean yellow dwarf virus (BeYDV). The expression system is preferably inducible to avoid interference with plant growth and development. Developmental cues, such as fruit ripening, are employed to trigger expression of the foreign protein using a tissue-specific promoter. A single, stably integrated expression cassette for foreign protein is replicated extrachromosomally in ripening fruit, forming hundreds of transcriptionally competent copies. Preferred plant hosts include tomato as a model system and soybean for production of large quantities of protein at high total protein levels.
    • 基于植物病毒遗传元件的基因扩增系统大大增加了植物中的外来蛋白质产量。 描述了使用农业实践生长的重组植物中更安全和更经济的疫苗和抗体生产系统。 高水平的表达系统使用植物重症病毒的复制过程,以豆黄矮病毒(BeYDV)为例。 表达系统优选可诱导以避免干扰植物生长和发育。 使用诸如果实成熟的发育提示来使用组织特异性启动子来引发外源蛋白的表达。 用于外来蛋白质的单一稳定整合的表达盒在成熟果实中被染色体外复制,形成数百个可转录的拷贝。 优选的植物宿主包括番茄作为模型系统,大豆用于生产高蛋白总量的大量蛋白质。