Based on BBa_K1995008 (BH), we added INP-N to the upstream of BH. Using INP-N, we anchored BH 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 BH 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 PH and other parts and get the following chart. (Fig. 5)
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25INCOMPATIBLE WITH RFCIllegal AgeI site found at 429
- 1000INCOMPATIBLE WITH RFCIllegal SapI.rc site found at 238