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In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells
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JoVE Journal Genetica
In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells

In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells

DOI:
10.3791/55832-v

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11:56 min

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September 28, 2017

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, , , ,
1Department of Environmental and Occupational Health, School of Public Health, NIEHS Center for Environmental Exposures and Disease, Environmental and Occupational Health Sciences Institute,Rutgers University, 2Veterinary Drugs & Biologics Division,Animal and Plant Quarantine Agency

Capitoli

  • 00:05Titolo
  • 00:53Concstruction of PER2 Promoter Driven Destabilized Luciferase Reporter Vector
  • 03:06Transforming Ligated Vector pGL[hPer2P/Luc2P/Neo] to E. coli
  • 05:28Transient Transfection of Cells with the Circadian Vector
  • 07:46Establishment of the In Vitro Bioluminescence Assay in Transiently Transfected MCF10A Cells
  • 08:52Results: Effects of Chemicls on the Cellular Biological Clock
  • 10:16Conclusion

Summary

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Traduzione automatica

An in vitro bioluminescence assay to determine cellular circadian rhythm in mammary epithelial cells is presented. This method utilizes mammalian cell reporter plasmids expressing destabilized luciferase under the control of the PERIOD 2 gene promoter. It can be adapted to other cell types to evaluate organ-specific effects on circadian rhythm.

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