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    • 6. 发明申请
    • SURFACE DISPLAY-BASED YEAST TWO HYBRID SCREENING SYSTEM
    • 基于表面显示的YEAST双混合筛选系统
    • WO2009088994A1
    • 2009-07-16
    • PCT/US2009/000046
    • 2009-01-05
    • LOS ALAMOS NATIONAL SECURITY, LLCCAI, HongCHEN, Jun
    • CAI, HongCHEN, Jun
    • C12N1/00
    • C12N15/1037
    • A surface display-based yeast two hybrid (sdY2H) system for rapid and large cDNA library scale protein interactome networking screening is provided. The sdY2H system utilizes a combination of surface display markers, highly efficient yeast mating resulting in high transformation efficiencies, and positive cell enrichment using a robust immunomagnetic affinity purification. In addition, the system of the invention can be combined with multiparameter fluorescence markers allowing further quantitative fluorescence analysis/sorting, enabling a significant reduction in the number of false positives. In some embodiments, the system uses liquid culture, rather than plate-based culture, resulting in substantially shortened cell culturing times that significantly reduce background fluorescence, allowing better discrimination of positive cells.
    • 提供了一种基于表面显示的酵母双杂交(sdY2H)系统,用于快速和大型cDNA文库规模蛋白质相互作用组合筛选。 sdY2H系统利用表面显示标记物的组合,高效酵母交配导致高转化效率,并使用强大的免疫磁性亲和纯化进行阳性细胞富集。 此外,本发明的系统可以与允许进一步定量荧光分析/分选的多参数荧光标记组合,从而能够显着减少误报数量。 在一些实施方案中,该系统使用液体培养而不是基于板的培养物,导致基本上缩短的细胞培养时间,显着降低背景荧光,允许更好地鉴别阳性细胞。