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  3. Protein Complementation Assay

Protein Complementation Assay蛋白质互补测定法

释义：

In the protein-fragment complementation assay, the proteins of interest ("Bait" and "Prey") are covalently linked at the genetic level to incomplete fragments of a third protein (known as the "reporter") and are expressed in vivo, Interaction between the "bait" and the "prey" proteins brings the fragments of the "reporter" protein in close enough proximity to allow them to reform and become the functional reporter protein. Typically enzymes which confer resistance to antibiotics, such as Dihydrofolate reductase or Beta-lactamase, or proteins that give colorimetric or fluorescent signals are used. The Bait protein is generally the protein under study and the methods are readily adaptable to highthroughput mode.

在蛋白质片段互补测定法中，感兴趣的蛋白质 (“诱饵”和“猎物”) 在基因层面上与第三种蛋白质 (称为“报告蛋白”) 的不完全片段共价连接，并在体内表达。“诱饵”和“猎物”蛋白之间的相互作用将报告蛋白的片段拉得足够近，从而使其重新组装并成为功能性的报告蛋白。通常使用能够赋予抗生素抗性 (如二氢叶酸还原酶或β-内酰胺酶) 的酶，或者能给出比色或荧光信号的蛋白质。诱饵蛋白通常是研究的蛋白质，这些方法可以轻松适应高通量模式。

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