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

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

DOI:
10.3791/50448-v

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11:40 min

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June 25, 2013

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, , ,
1Department of Biochemistry and Molecular Biology,State University of New York Upstate Medical University

Capitoli

  • 00:05Titolo
  • 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

Traduzione automatica

Traduzione automatica

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.

Tags

Keywords: Mgm101Recombinant ProteinMBP-taggedE. Coli ExpressionAffinity ChromatographyCationic Exchange ChromatographySize Exclusion ChromatographyMitochondrial DNA RepairRad52-type Recombination Proteins

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