Part:BBa_K4691000

recA-HRP-eGFP

SOS Response is a stress response of the organism itself, when bacterial DNA is damaged to a certain extent, it will repair the activity of related genes by activating the SOS response.In the genome of E. coli, many genes are found to participate in the damage repair process of SOS reaction, under normal circumstances, DNA damage repair gene activity is inhibited by LexA inhibitory protein, when SOS reaction occurs, RecA protein activity is activated, triggering the self-cleavage of LexA protein, and then initiating the transcriptional activity of downstream genes, expressing fluorescent proteins, observing bacterial growth to qualitatively evaluate biological damage effects, and using fluorescence signals to quantitatively evaluate DNA damage.

HrpRS and PhrpL are derived from the ultrasensitive pathogenic gene regulatory network (HRP) (HRP) in the gram-negative bacterium Pseudomonas lilac, which enables high-gain transcription amplification.Specifically, the combination of HrpR protein and HrpS protein forms an ultrasensitive complex that binds to the upstream active sequence of the dependent σ54 factor hrpL promoter PhrpL, and uses the energy generated by ATP hydrolysis to transform the inactive σ54-RNAP-hrpL transcription complex into an active complex, thereby initiating the expression of downstream proteins and achieving high-gain signal output.

We combined the recA promoter and the HRP amplifier to construct the recA-HRP-eGFP gene circuit in anticipation of a more sensitive DNA damage sensor

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 2010
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1220
  Illegal BsaI.rc site found at 2893
  Illegal SapI.rc site found at 1853