Part:BBa_K3945012:Design
Elektra 2: An electrochemical- measurement system for rare earth elements
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 89
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 89
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 89
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 89
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The reason histidine was chosen to be the amino acid that would covalently attach to the ruthenium molecule was because there were no other histidine amino acids in lanmodulin. This will allow for selective binding as it is very important that there are no other ruthenium molecules that are attached to lanM except on the end (as it could affect REE binding in addition to creating false positives in Elektra).
In order to find the best construct for Elektra, we ordered three constructs each with a differing number of cysteines on the c-terminal end (one cysteine - Elektra 1, two cysteines- Elektra 2, or three cysteines - Elektra 3). This will allow us to determine which one will give us the best protein density that will not create too much background signal, allow us to easily attach lanM onto the gold electrode, and be selective and sensitive to lanthanides.
Source
The sequence was obtained from the genome of Methylobacterium extorquens