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.
Original language | English (US) |
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Pages (from-to) | 63-73 |
Number of pages | 11 |
Journal | Methods in Molecular Biology |
Volume | 1263 |
DOIs | |
State | Published - 2015 |
Keywords
- Cell-free
- High-throughput screening
- Protein degradation
- Protein turnover
- Small molecules
- Xenopus egg extract
- Xenopus laevis
ASJC Scopus subject areas
- Molecular Biology
- Genetics