• Broadus MR, Yew PR, Hann SR, Lee E. Small-molecule high-throughput screening utilizing Xenopus egg extract. Methods in molecular biology (Clifton, N.J.). 1263. 63-73. PMID: 25618336 [PubMed]. PMCID: PMC4492114. NIHMSID: NIHMS703813.


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.