A Cell-Free System for Investigating Human MARF1 Endonuclease Activity
| Autor: | Marc R. Fabian, Hana Fakim |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
biology Oligonucleotide Chemistry Endoribonuclease activity Endoribonuclease RNA Cell-free system law.invention Exoribonucleases 03 medical and health sciences Endonuclease 0302 clinical medicine Biochemistry law Recombinant DNA biology.protein 030217 neurology & neurosurgery 030304 developmental biology |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071609347 |
| DOI: | 10.1007/978-1-0716-0935-4_21 |
| Popis: | Experiments in cell cultures have been useful for investigating a number of RNA endonucleases. However, endonuclease decay intermediates are often challenging to study in cellulo, as decay intermediates are rapidly degraded by exoribonucleases. Thus, cell-free assays have been critical for assessing endonuclease kinetics. Here, we describe such an in vitro assay to analyze endoribonuclease activity using recombinant proteins and end-radiolabeled RNA oligonucleotides. Specifically, we detail a protocol for assaying the endoribonuclease activity and kinetics of the human MARF1 protein. |
| Databáze: | OpenAIRE |
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