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Chapitres
- 00:05Titre
- 02:10Construction of MBP-Mgm101 Fusion Protein
- 03:16Expression, Induction, Cell Lysis and DNase I Treatment
- 05:28Purification on Amylose Column
- 07:07PreScission Protease Cleavage and Cation Exchange Chromatography
- 08:42Size Exclusion Chromatography
- 10:27Analysis of Purified Mgm101
- 11:23Conclusion
The yeast mitochondrial nucleoid protein, Mgm101, is a Rad52-type recombination protein that forms large oligomeric rings. A protocol is described to prepare soluble recombinant Mgm101 using the Maltose Binding Protein (MBP)-tagging strategy coupled with cation exchange and size exclusion chromatography.
