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

Capitoli

  • 00:04Titolo
  • 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

ThylakoidsProtein TransportChloroplast MembranesProtein TargetingPercoll GradientChloroplast IsolationPea ShootsHomogenizationCentrifugationChlorophyll Concentration

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