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    • 6. 发明申请
    • ISOLATION AND USE OF RYANODINE RECEPTORS
    • RYANODINE受体的分离和应用
    • WO2004027042A2
    • 2004-04-01
    • PCT/US2003/029834
    • 2003-09-23
    • E. I. DU PONT DE NEMOURS AND COMPANYCASPAR, TimothyCORDOVA, DanielGUTTERIDGE, StevenRAUH, James, J.SMITH, Rejane, M.WU, LihongTAO, Yong
    • CASPAR, TimothyCORDOVA, DanielGUTTERIDGE, StevenRAUH, James, J.SMITH, Rejane, M.WU, LihongTAO, Yong
    • C12N
    • C07K14/43563C07K14/705
    • The genes encoding ryanodine receptor homologs have been characterized from multiple insect families including lepidopteran tobacco budworm (Heliothis virescens), homopteran green peach aphid (Myzus persicae), corn plant hopper (Peregrinus maidis), cotton melon aphid (Aphis gossypii) and fruitfly (Drosophila melanogaster). The full-length genes have been isolated, cloned and amplified in bacterial cells. Expression in insect cells shows that the recombinant protein folds into a functional calcium release channel. The genes and their corresponding polypeptides have a number of uses including, but not limited to, the isolation of other pest ryanodine receptors, the development of screens to identify insecticidally active compounds, use of fragments of genes as pesticides, fragments of protein for antibody production, fragments of protein for determination of the structure of insecticide binding sites, and identification of insecticides that disrupt the calcium balance in cells through other messengers that interact with the receptor calcium release mechanism. Methods are outlined for overcoming toxic effects of expressing recombinant proteins in host cells.
    • 编码兰尼碱受体同系物的基因已经从多种昆虫科目表征,所述昆虫科包括鳞翅目烟夜蛾(Heliothis virescens),同翅目绿桃蚜(Myzus persicae),玉米植物漏斗(Peregrinus maidis),棉瓜蚜 (Aphis gossypii)和果蝇(果蝇(果蝇))。 已经在细菌细胞中分离,克隆和扩增全长基因。 在昆虫细胞中的表达显示重组蛋白折叠成功能性钙释放通道。 这些基因及其相应的多肽具有许多用途,包括但不限于分离其它害虫兰尼碱受体,开发鉴定杀虫活性化合物的筛选,使用基因片段作为农药,用于抗体产生的蛋白质片段 ,用于测定杀虫剂结合位点结构的蛋白质片段,以及通过与受体钙释放机制相互作用的其他信使破坏细胞中钙平衡的杀虫剂的鉴定。 概述了克服在宿主细胞中表达重组蛋白的毒性作用的方法。