Coding

Part:BBa_K925004

Designed by: Michael Paul Lockhart   Group: iGEM12_St_Andrews   (2012-09-24)

NiTagGST

This part consists of a glutathione S-transferase (GST) protein that has been modified to include a short peptide sequence allowing it to bind to nickel. The purified protein will bind to both GST and nickel, giving a double ended tag and a mechanism by which to recover nickel from solution.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 292
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 292
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 292
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 292
  • 1000
    COMPATIBLE WITH RFC[1000]


Description

This part combines the protein glutathione S-transferase (GST) with a novel nickel binding peptide. This provides a double ended tagging protein that binds efficiently to nickel.

Characterisation

Our NiTagGST protein was purified and was proven to bind to both GST and nickel beads, showing that both ends of the fusion protein were functional. A second characterisation technique that was used was UV-vis. This allowed us to see the shift in the λmax of a NiCl2 solution. The samples (with excess nickel removed) were sent to the University’s School of Geography and Geosciences for further conformation of presence of nickel using ICP-MS. We are currently awaiting the results of this.

Results

Our results show that the NiTagGST protein functions to bind to both nickel and to GST beads.

Conclusion

The BioBrickTM produced proves that it is possible to add short metal binding peptides to the end of GST protein while maintaining the function of both sequences.

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Categories
Parameters
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