Shuttle phasmid

Shuttle phasmids, sometimes called phagemids, are chimeric DNA vectors that replicate as plasmids in Escherichia coli as mycobacteriophages and in mycobacteria.[1]

Application

Shuttle phasmids, sometimes called phagemids,[2] allow the insertion of foreign genetic material into the mycobacterial genome through homologous recombination (HR) or transposons.[3]

Transformation efficiencies achieved through shuttle phasmids far exceed those resulting from electroporation.[4]

History

In 1987, William Jacobs Jr. et al. utilized a shuttle phasmid for the first successful introduction of foreign DNA into mycobacteria. The employed shuttle phasmid—a term they coined—consisted of an E. coli bacteriophage lambda cosmid inserted into a non-essential region of mycobacteriophage TM4.[5][4]

Bardarov et al first created temperature-sensitive shuttle phasmids that replicate and form plaques at 30°C but are unable to do so at 37°C.[4]

Bardarov et al also adapted shuttle phasmids to insert or knock out genes via homologous recombination, a process termed specialized transduction.[4]

Shuttle phasmids enabled the isolation of mc²155, the fist plasmid-transformable Mycobacterium smegmatis strain.[6]

References

Citations

  1. ^ Jacobs 2014, p. 1.
  2. ^ Lamrabet & Drancourt 2012, p. 368.
  3. ^ Chhotaray et al. 2018, p. 286.
  4. ^ a b c d Borgers et al. 2019, p. 3760.
  5. ^ Sparks et al. 2023, p. 3.
  6. ^ Sparks et al. 2023, p. 16.

Works cited

  • Borgers, Katlyn; Vandewalle, Kristof; Festjens, Nele; Callewaert, Nico (2019). "A Guide to Mycobacterium Mutagenesis". The FEBS Journal. 286 (19): 3757–3774. doi:10.1111/febs.15041. PMID 31419030.
  • Chhotaray, Chiranjibi; Tan, Yaoju; Mugweru, Julius; Islam, Md Mahmudul; Hameed, H.M. Adnan; Wang, Shuai; Lu, Zhili; Wang, Changwei; Li, Xinjie; Tan, Shouyong; Liu, Jianxion; Zhang, Tianyu (2018). "Advances in the development of molecular genetic tools for Mycobacterium tuberculosis". Journal of Genetics and Genomics. 45 (6): 281–297.
  • Jacobs, William (2014). "Gene Transfer in Mycobacterium tuberculosis: Shuttle Phasmids to Enlightenment". Microbiology Spectrum. 2 (2): 1–22. doi:10.1128/microbiolspec.MGM2-0037-2013. PMC 4725585. PMID 26105819.
  • Lamrabet, Otmane; Drancourt, Michel (2012). "Genetic Engineering of Mycobacterium tuberculosis: A Review". Tuberculosis. 92 (5): 365–376.
  • Sparks, Ian L.; Derbyshire, Keith M.; Jacobs, William R.; Moritaa, Yasu S. (2023). "Mycobacterium smegmatis: The Vanguard of Mycobacterial Research". Journal of Bacteriology. 205 (1): e0033722. doi:10.1128/jb.00337-22. PMC 9879119. PMID 36598232.

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