Difference between revisions of "Part:BBa K1189018"
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| − | <p><b>Figure | + | <p><b>Figure 2.</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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Revision as of 05:02, 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).
Figure 1. Ribbon visualization of a fully assembled ferritin protein. Figure 2. Image of the colours of ABTS and TMB (10 mg/mL for both) after reacting with Prussian blue ferritin. 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1289