In Vitro Reconstitution of the Drosophila melanogaster CCR4-NOT Complex to Assay Deadenylation
摘要
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.