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Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)
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Biologia
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JoVE Journal Biologia
Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

DOI:
10.3791/50042-v

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11:31 min

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March 15, 2013

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, ,
1Department of Biophysics and Biophysical Chemistry,Johns Hopkins University School of Medicine, 2Changchun Institute of Applied Chemistry,Chinese Academy of Sciences , 3Department of Physics,Jilin University

Capitoli

  • 00:05Titolo
  • 01:54Strain Engineering Workflow
  • 03:11Sample Preparation
  • 06:17Imaging and Acquisition of Timelapse Movies
  • 08:48Results: Typical Data from a CoTrAC Experiment
  • 11:06Conclusion

Summary

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We describe a fluorescence microscopy method, Co-Translational Activation by Cleavage (CoTrAC), to image the production of protein molecules in live cells with single-molecule precision without perturbing the protein's functionality. This method has been used to follow the stochastic expression dynamics of a transcription factor, the λ repressor CI 1.

Tags

Single-molecule ImagingGene RegulationIn VivoCotranslational Activation By Cleavage (CoTrAC)Fluorescence MicroscopyProtein ProductionLive CellsFluorescent ReporterMembrane TargetingProtease RecognitionProtein Of InterestFluorescence ImagingPhotobleachingCell LineagesGene ExpressionLambda Repressor CISystems Biology

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