Part:BBa_K1995002

PHG (INP-N-HG)

Based on BBa_K1995000 (BHG), we added INP-N to the upstream of BHG. Using INP-N, we anchored BHG out of the outer membrane of E.coli in order to increase the binding efficiency of cupric and mercuric ions.

Ice nucleation protein (INP)

To increase the binding efficiency and detect the concentration of heavy metals, we used INP to anchor BHG out of the outer membrane of E. coli.

Ice nucleation protein (INP) is an excretion surface protein. It is widely used for establishing germ surface display system. It contains an internal repeated domain (IRD) and an N-terminal anchoring domain (fig.1). Studies have shown that decrease the time of repetition of IRD can also get a better-off rate of displaying. So we decreased the time of repetition of IRD to reduce pressure of synthesis of E. coli.

Binding heavy metals

We detect the binding efficiency of PHG and other parts and get the following chart. (Fig. 2)

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 429
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 238