Part:BBa_K2588007

Peptidoglycan synthesis inhibitor mraZ

We designed mraZ in order to develop a stable culture of E.coli with inhibited growth. As a part of the Growth Inhibition Module mraZ disables the synthesis of peptidoglycan, the main part of the bacterial cell wall. Therefore, the cells are no longer able to expand due to their missing cell wall material. In combination with the other parts of the Growth Inhibition Module, the cells don’t divide, grow or replicate DNA anymore. That leads to a stable number of cells in the culture, which sustain themselves as long as they have all nutrients.

Usage and Biology

mraZ is a highly conserved transcriptional regulator ahead of the division and cell wall (dcw) cluster in E. coli.¹ By binding directly to the DNA, it negatively regulates its own expression and that of other genes in the proximal part of the dcw cluster.² It is not clear yet, whether it also regulates the expression of other genes.² Furthermore, overproduction of this protein is lethal and perturbs the division of cells by destabilizing the divisome but it does not affect the FtsZ levels or prevent the assembly of the Z ring.²
In our project, mraZ was used as a part of the growth inhibition module, BBa_K2588021.

Design considerations

To avoid unintentional restrictions, we had to change one base from G to A by using a modified primer. To avoid side effects or dysfunctions we made sure that the amino acid sequence stayed the same.

References

1. The Highly Conserved MraZ Protein Is a Transcriptional Regulator in Escherichia coli, 2014, 10.1128/JB.01370-13
2. https://www.uniprot.org/uniprot/P22186

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]