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化学感受器配体结合域的高效复用与纯化方法
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Biochemistry
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JoVE Journal Biochemistry
Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
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化学感受器配体结合域的高效复用与纯化方法

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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

提出了一种 dCACHE 周质配体结合域的复用过程, 该方法由包涵体化学感受器 Tlp3 和纯化, 产生毫克量的蛋白质.

Tags

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

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