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监测细胞自主的生物钟节律基因表达,利用荧光素酶生物发光记者
JoVE Journal
Biology
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JoVE Journal Biology
Monitoring Cell-autonomous Circadian Clock Rhythms of Gene Expression Using Luciferase Bioluminescence Reporters
DOI:

10:38 min

September 27, 2012

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Chapters

  • 00:05Title
  • 02:01Construction of Lentiviral Luciferase Reporters
  • 02:28Production of Lentiviral Particles: Seed Cells (Day 1)
  • 03:05Transient Transfection: CaPO4/DNA Precipitation (Day 2)
  • 04:23Harvest Viral Particles (Days 3-4)
  • 05:02Infection of 3T3 Cells (Day 3 Onward)
  • 06:12Synchronization and Change to Recording Medium
  • 07:02Bioluminescence Recording of Reporter Cells
  • 07:26Data Analysis and Presentation
  • 08:12Results: Phase-specific Circadian Reporters
  • 09:47Conclusion

Summary

Automatic Translation

生物钟单个细胞内发挥作用,也就是说,它们是细胞的自我。在这里,我们描述了使用非侵入性,实时荧光素酶为基础的生物发光技术产生的细胞自主的时钟模型的方法。记者细胞提供听话的,功能性的昼夜节律生物学研究的模型系统。

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