Part:BBa_K2842710

NikR with TerL-C intein

E. coli nickel binding protein, NikR, assembled into BBa_K2842669 (Intein Passenger).

NikR is a small protein (133 aa) with high affinity (6.8 pM) to nickel. It is a two-domain protein and the C-terminal alone is sufficient to bind nickel tightly (2.2 pM). [1,2] We used NikR as a prrof of concept for our standardised intein fictionalisation method by assembling it in a modular fashion from BBa_k2842669. This construct could be used to functionalise intein polymers developed from BBa_K2842680 and BBa_K2842690 with the ability to bind and sequester Nickle ions from solution.

Figure 1: colony PCR of functionalised Intein Passenger BBa_K2842669 <p> (1) NikR with TerL-C intein BBa_K2842710
(2) ER protein with TerL-C intein BBa_K2842720
*edited to show relevant bands

[1] Chivers PT, Sauer RT. NikR is a ribbon-helix-helix DNA-binding protein. Protein Science. 1999;8(11):2494-500. [2] Chivers PT, Sauer RT. NikR repressor: High-affinity nickel binding to the C-terminal domain regulates binding to operator DNA. Chemistry & Biology. 2002;9(10):1141-8.

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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 771
  Illegal BsaI.rc site found at 28