在体体外重建果蝇 CCR4-NOT 复合物以检测去腺苷酸化。
In Vitro Reconstitution of the Drosophila melanogaster CCR4-NOT Complex to Assay Deadenylation.
机构信息
Institute of Biochemistry and Biotechnology and Charles Tanford Protein Center, Martin Luther University Halle-Wittenberg, Halle, Germany.
RNA Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD, USA.
出版信息
Methods Mol Biol. 2024;2723:19-45. doi: 10.1007/978-1-0716-3481-3_2.
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.
CCR4-NOT 复合物是一种多亚基聚（A）特异性 3' 外切核酸酶，可催化 mRNA 的去腺苷酸化。在本章中，我们描述了表达和纯化重组果蝇 melanogaster CCR4-NOT 的程序。此外，我们还提供了制备放射性标记 RNA 底物和进行体外去腺苷酸化测定的方案。
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