Part:BBa_K925002

HisTagGST

This part produces glutathione S-transferase (GST) protein with a histidine tag attached to the end. The protein produced binds to nickel beads.

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 polyhistidine tag. This provides a double ended tagging protein and was used to prove that the concept of adding short metal binding peptides to the end of GST could produce a functional protein that would bind to metal as well as to GST beads.

Characterisation

Our HisTagGST 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 λ_max 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 HisTagGST protein functions to bind to both nickel and to GST beads.

Conclusions

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.