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Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
JoVE Journal
Biologia
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JoVE Journal Biologia
Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
DOI:

11:40 min

June 25, 2013

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Capítulos

  • 00:05Título
  • 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

Summary

Tadução automática

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.

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