Composite

Part:BBa_K1631010

Designed by: Yuto Yamanaka   Group: iGEM15_UT-Tokyo   (2015-09-14)

rbs - Colicin Lysis Protein - d.term

This part is a composite part of Translational unit of colicin lysis protein(BBa_K1631003) and double termnator (BBa_B0015).

Usage and Biology

Colicins are a cytotoxins which are released to environment and kill other related strains.

Release of colicin involves one protein; Colicin Lysis Protein. Colicin lysis protein allows colicins to be released. The mechanism how the colicin lysis protein allows colicin release has not been fully elucidated, but it is sure that this protein raise membrane permiability and cause quasilysis.[1] After Colicin released, they diffuse through the medium and bind to the receptor on the target cell membrane. Then, they are imported to the cytoplasm or cytoplasmic membrane of target cell by Tol-system or Ton-system. (If you want to know the mechanism of the Colicin import, see [1])
Colicin producing cells also express Colicin Immunity Protein in order to protect themselves from cytotoxity of colicin.
Colicins have variety cytotoxity such as DNase activity, RNase activity or Pore forming across inner membrane.

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We tried to confirm the quasi-lysis, but regardless of induction with 1mM IPTG, OD600 is decreased after about 4 hours. This maybe caused by the leak from lac promoter(BBa_R0011). It is considered that quasi-lysis is occuered at low expression level of Colicin Lysis Protein, and even if the expression level is bigger than that point, the effect to E. coli would not so change.

Reference
[1]Cascales, E., Buchanan, S. K., Duché, D., Kleanthous, C., Lloubes, R., Postle, K., ... & Cavard, D. (2007). Colicin biology. Microbiology and Molecular Biology Reviews, 71(1), 158-229.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 25
    Illegal AgeI site found at 136
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None