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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
JoVE Revista
Biología
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JoVE Revista Biología
Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters

Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters

DOI:
10.3791/51153-v

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09:22 min

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November 26, 2013

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, , ,
1The Lewis-Sigler Institute for Integrative Genomics,Princeton University, 2Department of Molecular Biology,Princeton University, 3Division of Chemistry and Chemical Engineering,California Institute of Technology

Capítulos

  • 00:05Título
  • 01:33Creation of Artificial Transcription Factors (ATFs)
  • 02:28Creation of ATF Plasmid Reporter
  • 03:57Quantify the Effect of ATF Induction on Gene Expression Using GFP Reporter
  • 05:33Quantify the Effect of the ATF Induction on Cell Growth
  • 06:42Create Inducible Genomic Alleles
  • 07:20Results: Analysis of GFP Induction by ATF-promoter Pairs
  • 08:39Conclusion

Summary

Traducción Automática

Traducción Automática

This protocol describes an experimental procedure for the rapid construction of artificial transcription factors (ATFs) with cognate GFP reporters and quantification of the ATFs ability to stimulate GFP expression via flow cytometry.

Tags

Synthetic BiologyTranscription FactorsGene ExpressionReporter SystemsGFPHomologous RecombinationArtificial Transcription FactorsConditional Gene Expression

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