Xu C W, Mendelsohn A R, Brent R
Department of Genetics, Harvard Medical School, and Department of Molecular Biology, Massachusetts General Hospital, 50 Blossom Street, Boston, MA 02114, USA.
Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12473-8. doi: 10.1073/pnas.94.23.12473.
Two-hybrid methods have augmented the classical genetic techniques biologists use to assign function to genes. Here, we describe construction of a two-bait interaction trap that uses yeast cells to register more complex protein relationships than those detected in existing two-hybrid systems. We show that such cells can identify bridge or connecting proteins and peptide aptamers that discriminate between closely related allelic variants. The protein relationships detected by these cells are analogous to classical genetic relationships, but lend themselves to systematic application to the products of entire genomes and combinatorial libraries. We show that, by performing logical operations on the phenotypic outputs of these complex cells and existing two-hybrid cells, we can make inferences about the topology and order of protein interactions. Finally, we show that cells that register such relationships can perform logical operations on protein inputs. Thus these cells will be useful for analysis of gene and allele function, and may also define a path for construction of biological computational devices.
双杂交方法拓展了生物学家用于确定基因功能的经典遗传技术。在此，我们描述了一种双诱饵相互作用陷阱的构建，该陷阱利用酵母细胞来记录比现有双杂交系统中检测到的更为复杂的蛋白质关系。我们表明，此类细胞能够识别桥梁蛋白或连接蛋白以及可区分密切相关等位基因变体的肽适配体。这些细胞检测到的蛋白质关系类似于经典遗传关系，但便于系统地应用于整个基因组和组合文库的产物。我们表明，通过对这些复杂细胞和现有双杂交细胞的表型输出进行逻辑运算，我们可以推断蛋白质相互作用的拓扑结构和顺序。最后，我们表明记录此类关系的细胞能够对蛋白质输入进行逻辑运算。因此，这些细胞将有助于分析基因和等位基因功能，还可能为构建生物计算设备指明一条道路。
