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Isolation of Physiologically Active Thylakoids and Their Use in Energy-Dependent Protein Transport Assays
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Bioquímica
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JoVE Journal Bioquímica
Isolation of Physiologically Active Thylakoids and Their Use in Energy-Dependent Protein Transport Assays

Isolation of Physiologically Active Thylakoids and Their Use in Energy-Dependent Protein Transport Assays

DOI:
10.3791/58393-v

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12:25 min

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September 28, 2018

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, , ,
1Department of Plant Biology,University of California – Davis

Capítulos

  • 00:04Título
  • 00:30Chloroplast Isolation and Quantification
  • 03:30Isolation of Thylakoids
  • 04:42Stromal Extract (SE) Recovery
  • 06:33Transport through the cpTat Pathway
  • 07:44Transport through the cpSec1 and cpSRP Pathways
  • 08:59Thermolysin Treatment
  • 10:16Results: Transport of Proteins Through cpTat and cpSec1 and Integration Through cpSRP Pathways
  • 11:18Conclusion

Summary

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We present protocols herein for high-yield isolation of physiologically active thylakoids and protein transport assays for the chloroplast twin arginine translocation (cpTat), secretory (cpSec1), and signal recognition particle (cpSRP) pathways.

Tags

Keywords: ThylakoidsProtein TransportChloroplast MembranesProtein TargetingPercoll GradientChloroplast IsolationPea ShootsHomogenizationCentrifugationChlorophyll Concentration

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