Part:BBa_K2129001

EutS compartment-forming protein

EutS is a single protein that forms bacterial microcompartments. Proteins tagged with EutC 1-19 amino acid sequence will localize inside of compartment.

Pictured: EutC-eGFP High (left) and EutS Medium with EutC-eGFP Low (right).

Note that not all cells will produce compartments, though many will.

Please see our wiki (CU-Boulder 2016) for the most complete information and discussion.

Usage and Biology

We have primarily used EutS in a several plasmid system with EutC-eGFP (BBa_K2129003). Best results for compartment formation are seen when using a medium expression level of EutS and a low level of EutC-eGFP. (Specifically, BBa_K314110 f1 phage origin with BBa_J23114 (Low) and BBa_J23110 (Medium) constitutive promoters.)

For our project, one of our plasmids was an Ampicillin resistance plasmid. We advise against using an ampicillin resistance plasmid in the future, as we suspect that this causes some cells to grow extremely long and therefore be somewhat harder to characterize.

Characterization Chart

Note that some data taken is not present on this chart, but it is representative of what you can expect with different expression levels.

CU Boulder iGEM 2017

The CU Boulder team in 2017 has continued optimization of visualization for EutS compartments and obtained new and more robust data, including from another lab on the other side of the world.

Our new FusionRed-LVA marker (BBa_K2249000) allows more clear and consistent visualization of compartment formation.

""

Additionally, iGEM 2017 Bielefeld-CiBiTec performed their own transformations and visualization of EutS with FusionRed.

""

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]