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

Article DOI: 10.3791/50042-v • 11:31 min • March 15th, 2013
March 15th, 2013 •
1, 1,2,3, 1
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

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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.

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Single-molecule Imaging Gene Regulation In Vivo Cotranslational Activation By Cleavage Fluorescence Microscopy Protein Production Live Cells Single-molecule Precision Functionality Counting Protein Molecules Time Windows Fluorescence Microscope Laser Excitation Power Density Fluorescent Reporter Membrane Targeting Sequence Yellow Fluorescent Protein Protease Recognition Sequence Translationally Fused Temperature-controlled Microscope Stage Photobleaching Fluorescence Images Analysis Fluorescent Spots Detection
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