Concstruction of PER2 Promoter Driven Destabilized Luciferase Reporter Vector
3:06
Transforming Ligated Vector pGL[hPer2P/Luc2P/Neo] to E. coli
5:28
Transient Transfection of Cells with the Circadian Vector
7:46
Establishment of the In Vitro Bioluminescence Assay in Transiently Transfected MCF10A Cells
8:52
Results: Effects of Chemicls on the Cellular Biological Clock
10:16
Conclusion
Transcripción
The overall goal of this in vitro bioluminescence assay is to determine the effects of different environmental circadian disruptors and enhancers on the cellular circadian rhythm of mammary epithelial cells from mammary glands at normal or disease
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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.