Difference between revisions of "Part:BBa K1189019:Design"
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Latest revision as of 07:09, 2 November 2013
Heavy chain human ferritin with his-tag and E-coil under the lacI inducible promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 230
Illegal AgeI site found at 905 - 1000COMPATIBLE WITH RFC[1000]
Design notes
This part was codon optimized for expression in E. coli K12.
There is evidence in the literature that ferritin nanoparticles composed entirely of heavy ferritin subunits are more soluble in E. coli, in comparison to ferritin that is entirely light subunits (Lee et al., 2002).
Variation of subunit composition in ferritin can influence the iron uptake dynamics of the nanoparticle. To more closely match the natural uptake of iron, the iGEM Calgary 2013 team built heavy/light subunit fusions, forming ferritin nanoparticles with 12 disubunits. This method ensures a one to one proportion of heavy and light subunits in ferritin nanoparticles. Such fusions have proven expressible in E. coil (Dehal et al, 2010). Additionally, the Calgary team used this method to reduce N-termini on ferritin by half, to reduce the number of proteins fused to it and eliminate possible steric hindrance in the 3D space around ferritin.
Source
The sequence for this part was inspired by P02794 [UniParc] and generated using commercial synthesis. We synthesized BBa_K1189019 with a promoter, strong RBS, and fusion to a protein of interest in the 2013 iGEM Calgary project. This part was plasmid switched to pSB1C3 and submitted to the Parts Registry.