Difference between revisions of "Part:BBa K1189018"
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<partinfo>BBa_K1189018 short</partinfo>
 
<partinfo>BBa_K1189018 short</partinfo>
  
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<p>This part was created by fusing the heavy chain and light chains (<partinfo>BBa_K1189024</partinfo> <partinfo>BBa_K1189025</partinfo>) of human ferritin together. It is expressed under the lacI promoter (<partinfo>BBa_J04500</partinfo>) and has a his-tag for protein purification. An E-coil (<partinfo>BBa_K1189011</partinfo>) is included in order to allow binding of parts containing the respective K-coil (<partinfo>BBa_K1189010</partinfo>). Characterization of this part was done primarily with commercially purchased ferritin. This ferritin is structurally very similar to our recombinant ferritin and does not differ in its chemical properties.  </p>
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<p>This part was created by fusing the heavy chain and light chains (<partinfo>BBa_K1189024</partinfo> <partinfo>BBa_K1189025</partinfo>) of human ferritin together. It is expressed under the lacI promoter (<partinfo>BBa_J04500</partinfo>) and has a his-tag for protein purification. An E-coil (<partinfo>BBa_K1189011</partinfo>) is included in order to allow binding of parts containing the respective K-coil (<partinfo>BBa_K1189010</partinfo>). Characterization of this part was done primarily with commercially purchased ferritin. This ferritin is structurally very similar to our recombinant ferritin and does not differ in its chemical properties (Figure 1).  </p>
 
<p>This construct can be used as a reporter through a modification of the iron core to form Prussian Blue (Figure 2). The resulting molecule can then catalyze the formation of radicals from hydrogen peroxide, which can then cause a colour change in substrates such as TMB or ABTS (Figure 3).</p>
 
<p>This construct can be used as a reporter through a modification of the iron core to form Prussian Blue (Figure 2). The resulting molecule can then catalyze the formation of radicals from hydrogen peroxide, which can then cause a colour change in substrates such as TMB or ABTS (Figure 3).</p>
 
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<img src="https://static.igem.org/mediawiki/2013/1/18/UCalgary2013TRFerritinrender2png.png" alt="Ferritin" width="600" height="600">
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<img src="https://static.igem.org/mediawiki/2013/1/18/UCalgary2013TRFerritinrender2png.png" alt="Ferritin" width="300" height="300">
 
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<p><b>Figure 1.</b> Ribbon visualization of a fully assembled ferritin protein.</p>
 
<p><b>Figure 1.</b> Ribbon visualization of a fully assembled ferritin protein.</p>
 
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https://static.igem.org/mediawiki/2013/4/40/UCalgary2013TRPrussianblueferritinsynthesispartspage.png
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<p><b>Figure 2.</b> Comparison image of commercial ferritin to Prussian blue ferritin after the synthesis reaction. The synthesis reaction took place over a 12 hour time period. </p>
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https://static.igem.org/mediawiki/2013/6/6c/UCalgary2013TRSubstratecolourpartspage.png
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<img src="https://static.igem.org/mediawiki/2013/a/a9/UCalgary2013TRSubstratecolour.png" alt="Substrate Colours" width="250" height="300">
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<b>Figure 3.</b> Image of the colours of ABTS and TMB (10 mg/mL for both) after reacting with Prussian blue ferritin.
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<p><b>Figure 14.</b> Image of the colours of ABTS and TMB (10 mg/mL for both) after reacting with Prussian blue ferritin.</p>
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<img src="https://static.igem.org/mediawiki/2013/c/c7/UCalgary2013TRPrussianblueferritinsynthesis.png" alt="Prussian Blue Synthesis" width="400" height="200">
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<figcaption>
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<p><b>Figure 3.</b> Comparison image of commercial ferritin to Prussian blue ferritin after the synthesis reaction. The synthesis reaction took place over a 12 hour time period. </p>
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</figure>
 
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Revision as of 04:34, 28 September 2013

Human ferritin di-subunit with E coil w/ LacI promoter

This part was created by fusing the heavy chain and light chains (BBa_K1189024 BBa_K1189025) of human ferritin together. It is expressed under the lacI promoter (BBa_J04500) and has a his-tag for protein purification. An E-coil (BBa_K1189011) is included in order to allow binding of parts containing the respective K-coil (BBa_K1189010). Characterization of this part was done primarily with commercially purchased ferritin. This ferritin is structurally very similar to our recombinant ferritin and does not differ in its chemical properties (Figure 1).

This construct can be used as a reporter through a modification of the iron core to form Prussian Blue (Figure 2). The resulting molecule can then catalyze the formation of radicals from hydrogen peroxide, which can then cause a colour change in substrates such as TMB or ABTS (Figure 3).


Ferritin

Figure 1. Ribbon visualization of a fully assembled ferritin protein.

Substrate Colours

Figure 14. Image of the colours of ABTS and TMB (10 mg/mL for both) after reacting with Prussian blue ferritin.

Prussian Blue Synthesis

Figure 3. Comparison image of commercial ferritin to Prussian blue ferritin after the synthesis reaction. The synthesis reaction took place over a 12 hour time period.



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.rc site found at 1289