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    • 3. 发明申请
    • TARGETED FUSION PROTEINS AND METHODS FOR THE CHARACTERIZATION OF CELLULAR MEMBRANE DOMAINS
    • 有针对性的融合蛋白和细胞膜结构域特征的方法
    • WO2003006606A2
    • 2003-01-23
    • PCT/US2002/021495
    • 2002-07-09
    • OKLAHOMA MEDICAL RESEARCH FOUNDATIONRODGERS, William
    • RODGERS, William
    • C12N
    • C12N9/1205C07K14/705C07K2319/00C07K2319/02C07K2319/033C07K2319/60G01N33/5041G01N33/92
    • Cell membranes containing glycolipid-enriched membrane (GEM) and non-glycolipid-enriched membrane (non-GEM) domains are targeted using fusion proteins that are anchored in the cell membrane. Fusion proteins to target GEM (or non-GEM) domains are comprised of a selected fluorescent polypeptide, a membrane-targeting sequence of p56 lck (or pp60 c-Src for non GEM domains) and a linker inserted between the polypeptide and the membrane targeting sequence. Localization of fusion proteins in GEM and non-GEM domains is assessed using techniques including confocal microscopy, fluorescence-based techniques, and membrane fractionation. Using these techniques, compounds are screened for their effect on GEM and non-GEM domains of live cells. These fusion proteins therefore represent useful tools for studying subcellular trafficking and the function of discrete compartments in the plasma membrane.
    • 含有富含脂肪糖的膜(GEM)和非糖脂富集膜(非GEM)结构域的细胞膜使用锚定在细胞膜中的融合蛋白进行靶向。 靶向GEM(或非GEM)结构域的融合蛋白由选择的荧光多肽,p56 的膜靶向序列(或非GEM结构域的pp60 )和插入多肽 和膜靶向序列。 使用包括共焦显微镜,基于荧光的技术和膜分级技术的技术来评估GEM和非GEM结构域中融合蛋白的定位。 使用这些技术,筛选化合物对活细胞的GEM和非GEM结构域的影响。 因此,这些融合蛋白是用于研究亚细胞运输和质膜中离散隔室功能的有用工具。