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An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos
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In Vitro Reconstitution of the Drosophila melanogaster CCR4-NOT Complex to Assay Deadenylation
Filip Pekovic1,2, Elmar Wahle3
1Institute of Biochemistry and Biotechnology and Charles Tanford Protein Center, Martin Luther University Halle-Wittenberg, Halle, Germany. filip.pekovic@nih.gov.
Methods in Molecular Biology (Clifton, N.J.)
|October 12, 2023
Abstract:
The CCR4-NOT complex is a multi-subunit poly(A)-specific 3' exoribonuclease that catalyzes the deadenylation of mRNA. In this chapter, we describe procedures to express and purify recombinant Drosophila melanogaster CCR4-NOT. Furthermore, we provide protocols for preparing radioactively labeled RNA substrates and conducting in vitro deadenylation assays.

