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膜蛋白共翻译插入预成型纳米盘
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Co-Translational Insertion of Membrane Proteins into Preformed Nanodiscs
DOI:

08:24 min

November 19, 2020

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Chapters

  • 00:05Introduction
  • 00:45S30 Ribosomal Protein Lysate Preparation
  • 02:17Membrane Scaffold Protein 1E3D1 (MSP1E3D1) Nanodisc Assembly
  • 03:243-mL Continuous Exchange Cell-Free (CECF) Reaction Setup
  • 04:1060-Microliter CECF Reaction Setup
  • 06:07Results: Representative Preformed Nanodisc-Modified Membrane Protein Characterization
  • 07:40Conclusion

Summary

Automatic Translation

共翻译插入到预先形成的纳米盘中,使得在定义的脂质环境中研究无细胞合成的膜蛋白成为可能,而无需与去垢剂接触。该协议描述了基本系统组分的制备以及提高表达效率和样品质量的关键参数。

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