Difference between revisions of "Part:BBa K2842710"
 
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E. coli nickel binding protein, NikR, assembled into BBa_K2842669 (Intein Passenger).
 
E. coli nickel binding protein, NikR, assembled into BBa_K2842669 (Intein Passenger).
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<p> 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]
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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.
  
 
[[File:T--UCL--IP-NikR and IP-ER gel.png|400px|thumb|left|
 
[[File:T--UCL--IP-NikR and IP-ER gel.png|400px|thumb|left|
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<p>
 
<p>
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(1) NikR with TerL-C intein [https://parts.igem.org/BBa_K2842710 BBa_K2842710]<br />
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(1) NikR with TerL-C intein [https://parts.igem.org/Part:BBa_K2842710 BBa_K2842710]<br />
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(2) ER protein with TerL-C intein [https://parts.igem.org/BBa_K2842720 BBa_K2842720]<br />
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(2) ER protein with TerL-C intein [https://parts.igem.org/Part:BBa_K2842720 BBa_K2842720]<br />
 
*edited to show relevant bands
 
*edited to show relevant bands
 
</p>''' ]]
 
</p>''' ]]
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[1] Chivers PT, Sauer RT. NikR is a ribbon-helix-helix DNA-binding protein. Protein Science. 1999;8(11):2494-500.
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[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.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 02:59, 18 October 2018


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