Small-molecule high-throughput screening utilizing Xenopus egg extract.
AUTHORS
Broadus
MRMatthew R ,
Yew
PR P Renee ,
Hann
SR Stephen R ,
Lee
E Ethan .
Methods in Molecular Biology . 2015 ; 1263().
63-73
- PMID: 25618336[PubMed].
- PMCID: PMC4492114.
- NIHMSID: NIHMS703813
ABSTRACT
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.