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活细胞钙成像相结合的siRNA介导的基因沉默标识CA 2 +泄漏通道内质网膜和监管机制
JoVE Journal
Biologia
This content is Free Access.
JoVE Journal Biologia
Live Cell Calcium Imaging Combined with siRNA Mediated Gene Silencing Identifies Ca2+ Leak Channels in the ER Membrane and their Regulatory Mechanisms
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活细胞钙成像相结合的siRNA介导的基因沉默标识CA 2 +泄漏通道内质网膜和监管机制

DOI:
10.3791/2730-v

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13:40 min

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July 07, 2011

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, , ,
1Medical Biochemistry and Molecular Biology,Saarland University, 2Experimental and Clinical Pharmacology and Toxicology,Saarland University

Capitoli

  • 00:05Titolo
  • 02:16Preparation of Stock Solutions
  • 03:03Gene Silencing in HeLa Cells
  • 05:39Complementation of HeLa Cells
  • 07:07Live Cell Calcium Imaging
  • 09:58Representative Calcium Imaging Results
  • 12:58Conclusion

Summary

Traduzione automatica

Traduzione automatica

内质网在蛋白质生物合成和钙稳态的关键作用。我们已经建立了一个实验系统，使我们能够解决的Ca2 +泄漏通道的作用，并表征其公认的监管机制。该系统涉及的siRNA介导的基因沉默和活细胞内Ca2 +成像。

Tags

Live Cell Calcium ImagingSiRNA Mediated Gene SilencingCa2+ Leak ChannelsER MembraneRegulatory MechanismsEndoplasmic ReticulumProtein BiogenesisCalcium SignallingHeterotrimeric Sec61 ComplexNascent PolypeptidesPuromycinER Luminal Protein BiPExperimental SystemCoding RegionUntranslated Region (UTR)SEC61A1 GeneComplementation AnalysisIRES-GFP Vector

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