Fluorescent reporter DS6A mycobacteriophages reveal unique variations in infectibility and phage production in mycobacteria

Oren Mayer, Paras Jain, Torin R. Weisbrod, Daniel Biro, Libby Ho, Deborah Jacobs-Sera, Graham F. Hatfull, William R. Jacobs

Research output: Contribution to journalArticlepeer-review

22 Scopus citations


Mycobacteriophage DS6A is unique among the more than 8,000 isolated mycobacteriophages due to its ability to form plaques exclusively on mycobacteria belonging to the Mycobacterium tuberculosis complex (MTBC). Speculation surrounding this specificity has led to unsupported assertions in published studies and patents that nontuberculous mycobacteria (NTM) are wholly resistant to DS6A infection. In this study, we identified two independent nonessential regions in the DS6A genome and replaced them with an mVenus-expressing plasmid to generate fluorescent reporter phages Fφ2GFP12 and Fφ2GFP13. We show that even though DS6A is able to form plaques only on MTBC bacteria, infection of various NTM results in mVenus expression in transduced cells. The efficiency of DS6A in delivering DNA varied between NTM species. Additionally, we saw a striking difference in the efficiency of DNA delivery between the closely related members of the Mycobacterium abscessus complex, M. abscessus and Mycobacterium massiliense. We also demonstrated that TM4 and DS6A, two phages that do not form plaques on M. massiliense, differ in their ability to deliver DNA, suggesting that there is a phage-specific restriction between mycobacterial species. Phylogenetic analysis reveals that the DS6A genome has a characteristically mosaic structure but provided few insights into the basis for the specificity for MTBC hosts. This study demonstrates that the inability of the MTBC-specific phage DS6A to form plaques on NTM is more complex than previously thought. Moreover, the DS6A-derived fluorophages provide important new tools for the study of mycobacterial biology.

Original languageEnglish (US)
Pages (from-to)3220-3232
Number of pages13
JournalJournal of Bacteriology
Issue number23
StatePublished - 2016

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology


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