Béranger F, Aresta S, de Gunzburg J, Camonis J
INSERM U-248, Institut Curie, 26 rue d'Ulm, 75231 Paris cedex 05, France.
Nucleic Acids Res. 1997 May 15;25(10):2035-6. doi: 10.1093/nar/25.10.2035.
Two-hybrid methods detect interactions between two proteins fused at the C-termini of, respectively, a DNA-binding domain and the activation domain of a transcriptional activator. Thus the N-terminus of none of these proteins is available for interaction. We have tested whether a bait protein with a reverted polarity (i.e. N-bait-LexA-C) is suitable for two-hybrid interaction. We show that such constructs give a specific interaction signal, and document two cases where the sensitivity is dramatically increased. Such constructs might lead to the identification of partners missed during classical two-hybrid screens.
双杂交方法检测分别融合在转录激活因子的DNA结合结构域和激活结构域C末端的两种蛋白质之间的相互作用。因此，这些蛋白质中没有一个的N末端可用于相互作用。我们测试了具有反向极性的诱饵蛋白（即N-诱饵-LexA-C）是否适用于双杂交相互作用。我们表明，这样的构建体给出了特异性相互作用信号，并记录了两例灵敏度显著提高的情况。这样的构建体可能会导致鉴定出在经典双杂交筛选中遗漏的相互作用蛋白。
