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Immunology and Infection

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JoVE Journal Immunology and Infection
Enrichment of Native and Recombinant Extracellular Vesicles of Mycobacteria
 

Enrichment of Native and Recombinant Extracellular Vesicles of Mycobacteria

Article DOI: 10.3791/65138-v • 06:38 min • December 8th, 2023
December 8th, 2023 •
1, 1, 1,2, 1,3, 1, 1, 1, 1,4, 1, 1
1Bacterial Pathogenesis Laboratory, Infectious Diseases and Immunology Group, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, 2Clinical Microbiology Division, CSIR-Indian Institute of Integrative Medicine, 3ICAR-Research Complex for Eastern Region, 4Public Health Research Institute, Rutgers University

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This protocol details the enrichment of native mycobacterial extracellular vesicles (mEVs) from axenic cultures of Mycobacterium smegmatis (Msm) and how mCherry (a red fluorescent reporter)-containing recombinant MsmEVs can be designed and enriched. Lastly, it verifies the novel approach with the enrichment of MsmEVs containing the EsxA protein of Mycobacterium tuberculosis.

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Keywords: Extracellular Vesicles Mycobacteria Enrichment Recombinant Vaccine Candidates Affinity-based Approaches Mycobacterium Smegmatis Sautons Media Tween 80 Mid-exponential Stage Centrifugation MCherry Fluorescent Proteins
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