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March 18, 2026Journal of BacteriologyOpen Access

Modular toolkit to facilitate molecular manipulations in mycobacteria

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Authors

AKAsis Kumar KhuntiaADAnkita DablaYKYogita Kapoor

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Overview

This work develops a versatile toolkit to enhance gene expression in mycobacteria, suggesting exciting possibilities for genetic studies.

Key Points

  • The aim is to create a flexible toolkit for molecular manipulation in mycobacteria, improving gene expression capabilities.
  • Developed 32 integrative and 8 episomal E. coli -mycobacterium shuttle vectors.
  • Designed vectors with N- or C-terminal tags for protein visualization and purification.
  • Validated the system by expressing RbpA in Mycobacterium smegmatis and examined protein localization.
  • Enabled co-expression of multiple sigma factors and assessed their localization.
  • Vectors confirmed successful expression of tagged proteins in mycobacteria.
  • Achieved distinct localization of Wag31 and FtsZ proteins.
  • Demonstrated four gene co-expression for enhanced visualization.
  • SBP-HA-S tag improved protein complex purification efficiency.

Cite This Study

Khuntia et al. (2026) studied this question.

synapsesocial.com/papers/69ba424e4e9516ffd37a2775https://doi.org/10.1128/jb.00474-25
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