Composite

Part:BBa_K1897013:Design

Designed by: Ang Shi Hui   Group: iGEM16_NUS_Singapore   (2016-10-10)


Listeriolysin O


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1482
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 90
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

There is a HA tag available for characterisation of the protein produced via western blot. Note that the stop codon for LLO CDS BBa_K1897012 is shifted to after the HA tag. Also, the transcriptional terminators rrnBT1 (from BBa_B0010) + BBa_B0012 was derived from BBa_B0015, with the first 8 base pairs removed.


Source

Listeriolysin O (LLO) is derived from Listeria monocytogenes and is initially obtained from BBa_K177026. However, after sequencing, the results obtained for sequencing of BBa_K177026 did not appear to match that given on the page. We went on to synthesize new LLO (for BBa_K1897013) with the results obtained from sequencing, and also added a HA tag.

References

Kayal, S., & Charbit, A. (2006). Listeriolysin O: a key protein of Listeria monocytogenes with multiple functions. FEMS microbiology reviews, 30(4), 514-529.

Vadia, S., Arnett, E., Haghighat, A. C., Wilson-Kubalek, E. M., Tweten, R. K., & Seveau, S. (2011). The pore-forming toxin listeriolysin O mediates a novel entry pathway of L. monocytogenes into human hepatocytes. PLoS Pathog, 7(11), e1002356.