title: Small-molecule high-throughput screening utilizing Xenopus egg extract.
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.
authors.affiliation: Department of Cell and Developmental Biology, Vanderbilt University Medical Center, 465 21st Avenue South, U-4213A Learned Lab/MRBIII, Nashville, TN, 37232-8240, USA.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Broadus
authors.forename: P Renee
authors.initials: PR
authors.lastname: Yew
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Hann
authors.forename: Ethan
authors.initials: E
authors.lastname: Lee
mesh_terms: D000818:Animals; D002474:Cell-Free System; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D066298:In Vitro Techniques; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D010063:Ovum / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D054852:Small Molecule Libraries; D014981:Xenopus
chemical_list: D011993:Recombinant Fusion Proteins; D054852:Small Molecule Libraries; D008156:Luciferases