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एक उपन्यास और सुलभ दृष्टिकोण: स्ट्रक्चरल HEK 293F निलंबन कोशिकाओं में जीवविज्ञान के लिए पुनः संयोजक प्रोटीन अभिव्यक्ति
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
Recombinant Protein Expression for Structural Biology in HEK 293F Suspension Cells: A Novel and Accessible Approach
DOI:

11:20 min

October 16, 2014

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Chapters

  • 00:05Title
  • 01:48Starter Cultures
  • 03:43Large-scale Culture/Transfection in 1 L Roller Bottles
  • 05:16Protein-complex Purification from Whole Cell Extract
  • 09:20Results: Expression and Purification of the HDAC1, SDS3, and Sin3A Ternary Complex
  • 10:35Conclusion

Summary

Automatic Translation

The expression of recombinant proteins by mammalian systems is becoming an attractive method for producing protein complexes for structural biology. Here we present a simple yet highly efficient expression system using suspension grown mammalian cells to purify protein complexes for structural studies.

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