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重组蛋白表达的结构生物学在HEK 293F悬浮细胞:一种新的和可访问方法
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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