Part:BBa_K4604035

piG_08 (ampProm_mazE)

BioBrick piG_08 consists of the Amp promoter, mazE and the rrnB terminator. The backbone we used in the experiments is pGGAselect.

Usage and Biology

Toxin-antitoxin systems (TA-systems) play a crucial role in plasmid stability for naturally occurring plasmids[1]. Usually, the toxin targets essential cellular functions and causes growth arrest or cell death, to which the antitoxin acts as a counterpart. Toxin and antitoxin exhibit differences in their stability and lifespan[2]. While the antitoxin has a shortened lifespan due to its sensitivity to degradation, the toxin has a longer lifespan and is more stable. If the plasmid that contains the TA-system is lost, the antitoxin is rapidly degraded and the toxin concentration increases, leading to cell death. Therefore, when first discovered, TA systems were called “addiction modules” that ensure plasmid retention. The labile MazE (antitoxin) acts as a neutralizer to the stable MazF (toxin), which acts as an endoribonuclease. When MazF is present freely in the cell it cuts cellular RNA which ultimately leads to cell death. This BioBrick can be used to neutralize mazF.

References

[1]Espah Borujeni, Amin et al. “Automated physics-based design of synthetic riboswitches from diverse RNA aptamers.” Nucleic Acids Research 44 (2015): 1 - 13.

[2]BRZOZOWSKA, Iwona; ZIELENKIEWICZ, Urszula. Regulation of toxin–antitoxin systems by proteolysis. Plasmid, 2013, 70. Jg., Nr. 1, S. 33-41.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 226
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 515