A generic protocol for the affinity-purification of native macromolecular complexes from poxvirus-infected cells.
| Autor: | Bartuli J; Department of Biochemistry and Cancer Therapy Research Center (CTRC), Theodor Boveri-Institute, University of Wuerzburg, Am Hubland, 97074 Wuerzburg, Germany., Lorenzi I; Department of Biochemistry and Cancer Therapy Research Center (CTRC), Theodor Boveri-Institute, University of Wuerzburg, Am Hubland, 97074 Wuerzburg, Germany., Backes S; Institute for Virology and Immunobiology, University of Wuerzburg, Versbacher Straße 7, 97078 Wuerzburg, Germany., Grimm C; Department of Biochemistry and Cancer Therapy Research Center (CTRC), Theodor Boveri-Institute, University of Wuerzburg, Am Hubland, 97074 Wuerzburg, Germany., Fischer U; Department of Biochemistry and Cancer Therapy Research Center (CTRC), Theodor Boveri-Institute, University of Wuerzburg, Am Hubland, 97074 Wuerzburg, Germany.; Helmholtz Institute for RNA-based Infection Research (HIRI), Helmholtz Centre for Infection Research (HZI), 97080 Wuerzburg, Germany. |
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| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2022 Jan 21; Vol. 3 (1), pp. 101116. Date of Electronic Publication: 2022 Jan 21 (Print Publication: 2022). |
| DOI: | 10.1016/j.xpro.2021.101116 |
| Abstrakt: | The functional and structural characterization of macromolecular complexes requires protocols for their native isolation. Here, we describe a protocol for this task based on the recombinant poxvirus Vaccinia expressing tagged proteins of interest in infected cells. Tagged proteins and their interactors can then be isolated via affinity chromatography. The procedure is illustrated for the Vaccinia virus encoded multi-subunit RNA polymerase. Our protocol also allows the expression and isolation of heterologous proteins and hence is suitable for a broader application. For complete details on the use and execution of this profile, please refer to Grimm et al. (2019). Competing Interests: The authors declare no competing interests. (© 2021 The Authors.) |
| Databáze: | MEDLINE |
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