Part:BBa_K1616005

Holin/endolysin reversed - RBS and double T7 terminator

Holin - Endolysin

Holin and endolysin are toxins; after a period of late-gene expression, holin, an inner permeabilizing protein creates holes in the membrane in order to permeabilize it and then endolysin, a muralytic enzyme traverses the holes creating by holin and degraded the wall cell. This complex induce cell lysis of bacteria. (Young & Bläsi, 1995)

Reverse sequence

Our kill switch was designed to work in a system like pDawn, a light-inducible expression system. However, this system works with reversed sequence. That is why we designed a simple reversed Biobrick Holin-Endolysin (HE). This part was inserted into native pDawn for characterization of the Biobrick. This composite part is composed of the reverse sequence of Holin and the reverse sequence of Endolysin.

Fig. 1: Holin - Endolysin construction

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 306
  Illegal AgeI site found at 376
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

All BioBrick parts used for assembling the composite part are compatible with the RFC10 Biobrick standard.

Source

BBa_K1616005 is a composite part with BBa_K1616006 (endolysin reversed), BBa_K1616007 (reversed), BBa_K1616011 (RBS reversed) and BBa_B0025 (double terminator T7 reversed).

References

Young, R., & Bläsi, U. (1995). Holins: Form and function in bacteriophage lysis. In FEMS Microbiology Reviews (Vol. 17, pp. 191–205). http://doi.org/10.1016/01686445(94)00079-4