Department of Chemistry, University of Toronto, Toronto, Canada; Molecular Biotechnology, University of Potsdam, Potsdam, Germany.
Molecular Biotechnology, University of Potsdam, Potsdam, Germany.
J Mol Biol. 2019 Jan 18;431(2):391-400. doi: 10.1016/j.jmb.2018.11.011. Epub 2018 Nov 15.
We have developed a genetic circuit in Escherichia coli that can be used to select for protein-protein interactions of different strengths by changing antibiotic concentrations in the media. The genetic circuit links protein-protein interaction strength to β-lactamase activity while simultaneously imposing tuneable positive and negative selection pressure for β-lactamase activity. Cells only survive if they express interacting proteins with affinities that fall within set high- and low-pass thresholds; i.e. the circuit therefore acts as a bandpass filter for protein-protein interactions. We show that the circuit can be used to recover protein-protein interactions of desired affinity from a mixed population with a range of affinities. The circuit can also be used to select for inhibitors of protein-protein interactions of defined strength.
我们在大肠杆菌中开发了一种遗传回路，可以通过改变培养基中的抗生素浓度来选择不同强度的蛋白质-蛋白质相互作用。该遗传回路将蛋白质-蛋白质相互作用强度与β-内酰胺酶活性联系起来，同时对β-内酰胺酶活性施加可调节的正选择和负选择压力。只有当细胞表达具有设定的高通和低通阈值范围内亲和力的相互作用蛋白时，细胞才能存活；即，该回路因此充当蛋白质-蛋白质相互作用的带通滤波器。我们表明，该回路可用于从具有一系列亲和力的混合群体中回收具有所需亲和力的蛋白质-蛋白质相互作用。该回路还可用于选择具有特定强度的蛋白质-蛋白质相互作用抑制剂。
