Part:BBa_K3487005

Non-specific endonuclease

This is a new part in the registry. After being expressed, RalR acts as a non-specific endonuclease that can cleave methylated and unmethylated DNA, leading to cell death. We put it as a poisonous protein into the suicide switch and introduce all the strains we use to prevent organisms from escaping.

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]

2020 SZPT-CHINA

Characterization

It is crucial to verify that the new part works in the strain. We still don’t know how much of the endonuclease concentration can kill the chassis cells, which determines whether the suicide switch can work. Therefore, we set up a solid plate medium to detect the expression of endonuclease and check our operating procedures.

Result

The expression of RalR was good under the positive and negative controls, and the colony situation in the corresponding area was significantly less than that of the empty vector control group.

pET28a(+), vector positive control ddH2O, negative control RalR, Non-specific endonuclease

References

Guo Y, Quiroga C, Chen Q, et al. RalR (a DNase) and RalA (a small RNA) form a type I toxin-antitoxin system in Escherichia coli. Nucleic Acids Res. 2014;42(10):6448-6462.