Part:BBa_K4656010

PRM-CI857-PR-phIF-MazE-PphIF-MazF

The temperator-sensitive lambda CI repressor protein has a CI857 mutation, and when the temperature is reduced to 30℃, CI857 converges to synthesize a dimer, binds to the promoter PR and inhibits it, resulting in suppression of downstream phIF expression. Because the phIF expression product can inhibit the promoter PphIF and inhibit the expression of the toxin MazF downstream of it. Therefore, when the temperature decreased, the inhibition ability of phIF to promoter PphIF was weakened or even lost, and MazF expressed a large number of toxic proteins, resulting in the death of E. coli. (MazF and MazE belong to the toxin-antitoxin suicide system.)

However, when the temperature increased to 37℃, CI857 repressor protein denaturated, CI repressor could no longer bind to promoter PR and play a repressor role, and a large number of downstream phIF protein inhibited promoter PphIF. Therefore, the expression of toxin MazF in the pathway was inhibited, and the MazE antitoxin protein at the end of the gene route was expressed instead. Eventually the E. coli survived.

Usage and Biology

The gene route is temperature-controlled and can be used as a safety guarantee so that the engineered bacteria can die when they leave the human body (the temperature drops) and survive in the human body (37℃). It can solve the problem of handling engineered bacteria after they are discharged from the human body, ensuring that they play a role in the human body while doing not damage the environment.

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 1625
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]