Purification of Recombinant Human PARG and Activity Assays
| Autor: | Françoise Dantzer, Éléa Héberlé, Barbara Camuzeaux, Jean-Christophe Amé, Valérie Schreiber |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification PARG Lysis Protease Chemistry medicine.medical_treatment 010402 general chemistry medicine.disease_cause 01 natural sciences 3. Good health 0104 chemical sciences law.invention Sepharose 03 medical and health sciences 030104 developmental biology Enzyme Biochemistry law medicine Recombinant DNA Escherichia coli Poly(ADP-ribose) glycohydrolase |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493969920 |
| Popis: | The purification of Poly(ADP-ribose) glycohydrolase (PARG) from overexpressing bacteria Escherichia coli is described here to a fast and reproducible one chromatographic step protocol. After cell lysis, GST-PARG-fusion proteins from the crude extract are affinity purified by a Glutathione 4B Sepharose chromatographic step. The PARG proteins are then freed from their GST-fusion by overnight enzymatic cleavage using the preScission protease. As described in the protocol, more than 500 μg of highly active human PARG can be obtained from 1.5 L of E. coli culture. |
| Databáze: | OpenAIRE |
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