Difference between revisions of "Part:BBa K1189019"

Revision as of 00:32, 1 October 2013

Heavy chain human ferritin with his-tag and E-coil under the lacI inducible promoter

This heavy ferritin chain comes from humans. This part along with light ferritin (BBa_K1189024), form the ferritin nanoparticle, an iron-storage particle made up of 24 subunits. The formed nanoparticle is highly robust, remaining stable at extreme pHs and temperatures. The difference between light ferritin is that this chain contains a ferroxidase centre.

This nanoparticle can also be used as a reporter when the iron core is modified with potassium ferrocyanide to form Prussian Blue. The Prussian Blue ferritin can then act as a peroxidase mimic, similar to horseradish peroxidase, resulting in colour changes in the presence of hydrogen peroxide, and TMB or ABTS.

We added the lacI promoter (BBa_J04500), double terminator (BBa_B0015) and a his-tag in order for us to induce protein expression as well as purify it.

Applications of BBa_K1189019

This protein was successfully expressed and purified through the use of his-tags. WESTERN BLOT STUFF

We then modified the iron core in order to convert the protein into a peroxidase-like catalyst. Initial colourimetric testing was conducted and it was shown to be effective as the catalyst. PICTURE Qualitative kinetics testing was also conducted for commercial horsespleen ferritin to determine pH and temperature optimums, as well as Michaelis-Menten kinetics for the TMB substrates. PICTURESSSSSs

Sequence and Features