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Methods Mol Biol. 2015;1263:63-73. doi: 10.1007/978-1-4939-2269-7_5.

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

Author information

1
Department of Cell and Developmental Biology, Vanderbilt University Medical Center, 465 21st Avenue South, U-4213A Learned Lab/MRBIII, Nashville, TN, 37232-8240, USA.

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.

PMID:
25618336
PMCID:
PMC4492114
DOI:
10.1007/978-1-4939-2269-7_5
[Indexed for MEDLINE]
Free PMC Article

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