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化学感受器配体结合域的高效复用与纯化方法

JoVE Journal
Biochimica

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JoVE Journal Biochimica

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

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化学感受器配体结合域的高效复用与纯化方法

DOI:

10.3791/57092-v

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14:25 min

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December 12, 2017

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Mayra A. Machuca², Anna Roujeinikova^2,3

¹Infection and Immunity Program,Monash Biomedicine Discovery Institute, ²Department of Microbiology,Monash University, ³Department of Biochemistry and Molecular Biology,Monash University

Capitoli

00:05Titolo
00:46Expression of His₆-TIp3-LBD in E. coli
02:29Isolation and Denaturation of Inclusion Bodies
05:34Protein Refolding
07:39Purification of His₆-tagged Protein Using Immobilized Metal Ion Affinity Chromatogrphy
08:49His₆-tag Removal Using TEV Protease and Size-excluson Chromatography of Tlp3-LBD
11:06Results: Expression, Refolding, and Purification of Tlp3-LBD
12:41Conclusion

Summary

Traduzione automatica

提出了一种 dCACHE 周质配体结合域的复用过程, 该方法由包涵体化学感受器 Tlp3 和纯化, 产生毫克量的蛋白质.

Tags

Keywords: Chemoreceptor Ligand Binding Domain Refolding Purification Inclusion Bodies E. Coli Protein Expression IPTG Induction Cell Lysis Homogenization Centrifugation SDS-PAGE Buffer Exchange Solubilization Protein Purification

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