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Small-molecule high-throughput screening utilizing Xenopus egg extract.


Broadus MRMatthew R , Yew PR P Renee , Hann SR Stephen R , Lee E Ethan . Methods in Molecular Biology . 2015 ; 1263(). 63-73


Screens for small-molecule modulators of biological pathways typically utilize cultured cell lines, purified proteins, or, recently, model organisms (e.g., zebrafish, Drosophila, C. elegans). Herein, we describe a method for using Xenopus laevis egg extract, a biologically active and highly tractable cell-free system that recapitulates a legion of complex chemical reactions found in intact cells. Specifically, we focus on the use of a luciferase-based fusion system to identify small-molecule modulators that affect protein turnover.