Part:BBa_K4760000

ClyA-PbrR-His6

Pb-binding protein, PbrR, is diplayed on the outer membrane by ClyA(also called HlyE, see BBa_K257002).Secreted OMVs with this will possess Pb(II)-binding acitivity. On the C terminus of PbrR we linked a 6 aa His-Tag, which can faciliate the recovery of the OMVs through nickel.

Biology

The 34 kDa ClyA is enriched on the OMVs and structurally stretched outwards, making it a popular surface display scaffold on Gram- bactera OMVs. In our project, we used it to display the Cupriavidus metallidurans CH34 originated Pb(II)-binding protein PbrR, to test our idea of OMV-empowered bioremediation. We further added a His6 tag onto the C-terminus of the protein, through which we aimed to recover the OMVs with the binding of nichel and His tag. This did not prove working well in this part, yet was verified in our improved version.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 85
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]