Part:BBa_K1189018

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, which is structurally very similar to this recombinant ferritin (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).

This protein is a highly robust protein, remaining stable under extreme pH, temperature, and denaturing conditions. It is also highly accepting of fusino proteins, as it continues to form the nanoparticle despite fusions to both N-terminus and C-terminus. In addition, proteins fused to this protein have been found to be stabilized due to the fusion.

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

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