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用于无细胞重建的重组隔肽复合物的纯化和质量控制

JoVE Journal
Biochimica

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JoVE Journal Biochimica

Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution

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用于无细胞重建的重组隔肽复合物的纯化和质量控制

DOI:

10.3791/63871-v

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11:50 min

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June 23, 2022

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Gerard Castro-Linares, Jeffrey den Haan, Francois Iv, Carla Silva Martins, Aurélie Bertin, Manos Mavrakis, Gijsje H. Koenderink

¹Department of Bionanoscience, Kavli Institute of Nanoscience Delft,Delft University of Technology, ²Institut Fresnel, CNRS UMR7249,Aix Marseille Univ, Centrale Marseille, ³Institut Curie, Université PSL,Sorbonne Université

Capitoli

00:05Introduction
00:47Purification of Septin Hetero-Oligomers
04:29Quality Control of Purity and Integrity of the Septin Hetero-Oligomer
07:34Septin Functional Quality Control via Polymerization Analysis
08:57Results: Analysis of Purified Septin Quality
11:01Conclusion

Summary

Traduzione automatica

细胞骨架蛋白的体外重建是了解这些蛋白质基本功能特性的重要工具。本文描述了一种纯化和评估重组隔膜复合物质量的方案，重组隔膜复合物在细胞分裂和迁移中起核心作用。

Tags

Keywords: Recombinant Septin Complexes Cell-free Reconstitution Purification Quality Control Affinity Chromatography His-tagged Septins Strep-II Tagged Proteins FPLC Bacterial Expression IPTG Induction Cell Lysis Sonication Centrifugation Optical Density MsfGFP MEGFP Protein Degradation

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