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

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

DOI:
10.3791/61731-v

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08:34 min

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February 23, 2021

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

Chapitres

  • 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

Traduction automatique

Traduction automatique

MAPS technology has been developed to scrutinize the targetome of a specific regulatory RNA in vivo. The sRNA of interest is tagged with a MS2 aptamer enabling the co-purification of its RNA partners and their identification by RNA sequencing. This modified protocol is particularly suited for Gram-positive bacteria. 

Tags

Keywords: MS2-affinity PurificationRNA SequencingMAPSGram-positive BacteriaSRNARegulatory NetworksVirulence FactorsBiofilm FormationStaphylococcal InfectionsCell LysisAmylose ResinMBP-MS2 ProteinFlow-through Fraction

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