Difference between revisions of "Part:BBa K2916012"

 
 
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Nucleotide diphosphate kinase is a protein that is used in the PURE and OnePot PURE cell-free systems.
  
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===Usage and Biology===
 
===Usage and Biology===
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NDK plays a big part in the energy-recycling system. It catalyzes the reactions of phosphorylation of a histidine residue that produces NTP, which then initiates a chain of reactions in the system. Analyzing those reactions we can conclude that NDK is involved in the synthesis of nucleoside triphosphates (NTP), such as guanosine triphosphate (GTP), cytidine triphosphate (CTP) and uridine triphosphate (UTP) that are all parts of our energy solution.
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In our part, besides the sequence encoding for the protein we also have a hexahistidine-tag to allow us purify the protein.
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In our project we used NDK as a part of the protein solution needed for <html><a style="padding: 0px; margin: 0px;" href="https://2019.igem.org/Team:EPFL/OnePot_Pure"> OnePot PURE cell-free system </a></html> produced with the method of gravity flow affinity chromatography, as described in the <html><a style="padding: 0px; margin: 0px;" href="https://www.protocols.io/view/protein-purification-for-onepot-pure-cell-free-sys-8auhsew"> protocol </a></html> we designed.
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===Characterization===
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All the data regarding the characterization of that part can be found in the composite part: <html><a style="padding: 0px; margin: 0px;" href="https://parts.igem.org/Part:BBa_K2916047"> BBa_K2916047 </a></html>.
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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===Functional Parameters===
 
===Functional Parameters===
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Latest revision as of 03:30, 22 October 2019


NDK protein eqquiped with a 6x HIS affinity tag

Nucleotide diphosphate kinase is a protein that is used in the PURE and OnePot PURE cell-free systems.

Usage and Biology

NDK plays a big part in the energy-recycling system. It catalyzes the reactions of phosphorylation of a histidine residue that produces NTP, which then initiates a chain of reactions in the system. Analyzing those reactions we can conclude that NDK is involved in the synthesis of nucleoside triphosphates (NTP), such as guanosine triphosphate (GTP), cytidine triphosphate (CTP) and uridine triphosphate (UTP) that are all parts of our energy solution.

In our part, besides the sequence encoding for the protein we also have a hexahistidine-tag to allow us purify the protein.


In our project we used NDK as a part of the protein solution needed for OnePot PURE cell-free system produced with the method of gravity flow affinity chromatography, as described in the protocol we designed.



Characterization

All the data regarding the characterization of that part can be found in the composite part: BBa_K2916047 .

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 31
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]