Maier Richard H, Maier Christina J, Onder Kamil
Division of Molecular Dermatology, Department of Dermatology, Paracelsus Private Medical University Salzburg, Salzburg, Austria.
Methods Mol Biol. 2011;729:71-84. doi: 10.1007/978-1-61779-065-2_5.
The Yeast Two-Hybrid (Y2H) system is the most frequently used method for identifying protein-protein interactions. The use of recombination-amenable Y2H vectors would reduce time and effort for cloning prey or bait vectors, and increase the quality of Y2H screenings due to the production of improved screening libraries. These libraries can heighten the amount of new candidates in Y2H screenings significantly by representing more correct candidate genes in frame and outperform a classical Y2H library. The described vectors can be used for the construction of genomic, peptide, or cDNA-based Y2H libraries. Furthermore, the compatibility to newer ORFeome libraries is also given. Here, we describe a vector system for site-specific recombination and for the construction of high-content Y2H libraries. In summary, we will describe the construction of these vectors and the production of Y2H screening libraries.
酵母双杂交（Y2H）系统是用于鉴定蛋白质-蛋白质相互作用最常用的方法。使用易于重组的Y2H载体将减少克隆猎物或诱饵载体的时间和精力，并且由于产生了改进的筛选文库而提高了Y2H筛选的质量。这些文库通过在框架内代表更多正确的候选基因，可以显著增加Y2H筛选中新候选物的数量，并且优于经典的Y2H文库。所描述的载体可用于构建基于基因组、肽或cDNA的Y2H文库。此外，还具有与更新的ORFeome文库的兼容性。在这里，我们描述了一种用于位点特异性重组和构建高含量Y2H文库的载体系统。总之，我们将描述这些载体的构建以及Y2H筛选文库的产生。
