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MS2亲和力纯化与革兰阳性细菌中的RNA测序相结合

JoVE Journal
Biochemistry

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JoVE Journal Biochemistry

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria

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MS2亲和力纯化与革兰阳性细菌中的RNA测序相结合

DOI:

10.3791/61731-v

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08:34 min

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February 23, 2021

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Noémie Mercier, Karine Prévost, Eric Massé, Pascale Romby, Isabelle Caldelari, David Lalaouna

¹Université de Strasbourg, CNRS, ²Department of Biochemistry,Université de Sherbrooke

Chapters

00:05Introduction
01:05Bacteria Harvesting and Lysis
02:52MS2-affinity Purification
03:55RNA Extraction of Collected Fractions (CE, FT, W and E)
05:24Results: Constructs Validation and MAPS Controls
07:49Conclusion

Summary

Automatic Translation

MAPS 技术已开发出来，以仔细审查体内特定监管 RNA 的目标。感兴趣的 sRNA 标有 MS2 贴录器，通过 RNA 测序实现其 RNA 合作伙伴的共同纯化及其标识。此修改后的协议特别适合革兰氏阳性细菌。

Tags

MS2-affinity Purification RNA Sequencing MAPS Gram-positive Bacteria SRNA Regulatory Networks Virulence Factors Biofilm Formation Staphylococcal Infections Cell Lysis Amylose Resin MBP-MS2 Protein Flow-through Fraction

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