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Journal
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Biochemistry
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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
Please note that all translations are automatically generated.
Click here for the English version.
膜蛋白共翻译插入预成型纳米盘
DOI:
10.3791/61844-v
•
08:24 min
•
November 19, 2020
•
Roman Levin
,
Zoe Koeck
,
Volker Dötsch
,
Frank Bernhard
1
Institute of Biophysical Chemistry
,
Goethe University Frankfurt
Chapters
00:05
Introduction
00:45
S30 Ribosomal Protein Lysate Preparation
02:17
Membrane Scaffold Protein 1E3D1 (MSP1E3D1) Nanodisc Assembly
03:24
3-mL Continuous Exchange Cell-Free (CECF) Reaction Setup
04:10
60-Microliter CECF Reaction Setup
06:07
Results: Representative Preformed Nanodisc-Modified Membrane Protein Characterization
07:40
Conclusion
Summary
Automatic Translation
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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。
Tags
Co-translational Insertion
Membrane Proteins
Nanodisc
Detergent-free
Protein Folding
Lipid Effects
Cell-free Expression
Membrane Scaffold Protein
Cell Disruption
Dialysis
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