Part:BBa_K4156083

φ174E

φ174E is a kind of phage cleavage gene. Expression of this gene triggers bacterial lysis and death ^[1-3], thereby releasing therapeutic factors stored in the bacteria.

Usage and Biology

φ174E induces bacteria lysis in our constructed effector strains, thereby releasing intracellularly expressed therapeutic proteins.

Demonstration of the function of the lysis gene

Group: iGEM Team OUC-China 2023

Author: Qingyu Wu

Summary: Successfully demonstrated the lysis function of this gene

After induction with IPTG for 2-3 hours, the lysin protein can almost completely kill all bacteria. According to the results of SDS-PAGE in the figure below, it can be observed that there are almost no protein bands in the induced group. This is because a large amount of protease is released after cell death, leading to protein degradation, indicating that the lysin protein is functioning.

Fig 1 (A) Gene circuit (B) Colony PCR (C) SDS-PAGE M: Protein Marker 1: 0mM IPTG 2: 0.2mM IPTG

Characterization

We used φ174E in the E. coli lysis experiment with validation of therapeutic efficacy, and characterization could be seen at BBa_K4156100  ; BBa_K4156105 ； BBa_K4156109 .

References

1 Prindle A, Samayoa P, Razinkov I, Danino T, Tsimring LS, Hasty J. A sensing array of radically coupled genetic 'biopixels'. Nature. Dec 18 2011;481(7379):39-44. doi:10.1038/nature10722

2 Young KD, Young R. Lytic action of cloned phi X174 gene E. J Virol. Dec 1982;44(3):993-1002. doi:10.1128/jvi.44.3.993-1002.1982

3 Marguet P, Tanouchi Y, Spitz E, Smith C, You L. Oscillations by minimal bacterial suicide circuits reveal hidden facets of host-circuit physiology. PLoS One. Jul 30 2010;5(7):e11909. doi:10.1371/journal.pone.0011909

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