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Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
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Bioquímica
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JoVE Journal Bioquímica
Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

DOI:
10.3791/57092-v

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14:25 min

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December 12, 2017

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,
1Infection and Immunity Program,Monash Biomedicine Discovery Institute, 2Department of Microbiology,Monash University, 3Department of Biochemistry and Molecular Biology,Monash University

Capítulos

  • 00:05Título
  • 00:46Expression of His6-TIp3-LBD in E. coli
  • 02:29Isolation and Denaturation of Inclusion Bodies
  • 05:34Protein Refolding
  • 07:39Purification of His6-tagged Protein Using Immobilized Metal Ion Affinity Chromatogrphy
  • 08:49His6-tag Removal Using TEV Protease and Size-excluson Chromatography of Tlp3-LBD
  • 11:06Results: Expression, Refolding, and Purification of Tlp3-LBD
  • 12:41Conclusion

Summary

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A procedure is presented for the refolding of the dCACHE periplasmic ligand binding domain of Campylobacter jejuni chemoreceptor Tlp3 from inclusion bodies and the purification to yield milligram quantities of protein.

Tags

ChemoreceptorLigand Binding DomainRefoldingPurificationInclusion BodiesE. ColiProtein ExpressionIPTG InductionCell LysisHomogenizationCentrifugationSDS-PAGEBuffer ExchangeSolubilizationProtein Purification

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