Expression and Purification of pAG-MNase for CUT&RUN.
| Autor: | Kawakatsu T; Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan. riverwin@naro.affrc.go.jp. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2025; Vol. 2869, pp. 147-155. |
| DOI: | 10.1007/978-1-0716-4204-7_15 |
| Abstrakt: | Cleavage Under Targets and Release Using Nuclease (CUT&RUN) is a chromatin profiling strategy that releases specific DNA-protein complexes from in situ chromatin to the supernatant. CUT&RUN employs the hybrid protein pAG-MNase, composed of IgG binding proteins A and G, followed by micrococcal nuclease. Here, I describe the expression and purification of recombinant pAG-MNase as well as the assessment of its enzymatic activity. Purified pAG-MNase is hyperactive and enables large-scale analyses. (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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