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Direkte Protein Delivery for pattedyrsceller Brug Cell-permeable Cys<sub> 2</sub> -His<sub> 2</sub> Zink-finger Domæner
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JoVE Journal Biologia
Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys2-His2 Zinc-finger Domains

Direkte Protein Delivery for pattedyrsceller Brug Cell-permeable Cys<sub> 2</sub> -His<sub> 2</sub> Zink-finger Domæner

DOI:
10.3791/52814-v

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11:24 min

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March 25, 2015

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,
1Departments of Chemistry and Cell and Molecular Biology,The Scripps Research Institute, 2Shanghai Institute for Advanced Immunochemical Studies,ShanghaiTech University

Capítulos

  • 00:05Título
  • 01:23Cloning Emerald GFP into an Expression Vector Containing Two-finger ZiF Domains
  • 04:22Purification of the ZiF-EmGFP Fusion Protein
  • 07:17Protein Transduction of HeLa Cells
  • 10:07Results: Fusion of 2F-ZiF Domains to EmGFP Enables Delivery of EmGFP to HeLa Cells
  • 10:57Conclusion

Summary

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Zinkfinger-domæner er uløseligt celle-permeable og i stand til at mediere protein levering i en bred række af mammale celletyper. Her er en detaljeret trin-for-trin-protokollen til gennemførelse zink-finger-teknologi til intracellulær protein levering fremlagt.

Tags

Zinc-fingerProtein DeliveryCell-permeableCys2-His2Genome EngineeringIntracellular DeliveryCell MembraneProtein TransductionEnzyme ActivityCytosolic DeliveryMammalian Cells

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