Part:BBa_K4463002

SUMO-CsgA-SmtA
SmtA is a protein of Metallothionein family, which can bind to multiple metal ions, such as Cu(II), Cd(II) and Cu(II). CsgA is the major curli subunit protein of E.coli. Therefor, the surface display of SmtA is achieved by linking to CsgA. SUMO tag was added to improve the solubility of recombinant protein, CsgA and SmtA. Improved parts is based on part BBa_K519010.

In the IGEM project of XJTLU_China 2022, we added SUMO before SmtA to improve its function. We measured the amounts of binding Cu(II) and Cd(II), the results showed the function of SmtA performed better in the presence of SUMO.
We cultured modified E.coli in 1.8mM CuSO4 and 300μM CdCl2 respectively for 8 hours. Then we used ICP-MS to measure the concentration of metal ions adsorbed by modified E.coli. For more information of our measurement and result analysis, please visit our Experiments and Results.

Figure1. The left column is the concentration of Cu(II) adsorbed by control group that E.coli containing empty plasmid. The middle column is Cu(II) adsorbed by E.coli expressing SmtA. The right column is Cu(II) adsorbed by E.coli expressing SUMO-SmtA, which is also the part in this page. All groups grew in 1.8mM CuSO4 solution.

Figure2. The left column is the concentration of Cd(II) adsorbed by control group that E.coli containing empty plasmid. The middle column is Cd(II) adsorbed by E.coli expressing SmtA. The right column is Cd(II) adsorbed by E.coli expressing SUMO-SmtA, which is also the part in this page. All groups grew in 300μM CdCl2 solution.

The result showed that our improvement on SmtA is successful!

 

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 863
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 863
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 297
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 863
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 863
  Illegal AgeI site found at 891
  
• 1000
  COMPATIBLE WITH RFC[1000]