Part:BBa_K1223006:Design
His-Tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The His-Tag (or 6XHis) is a widely use sequence both as a reporter (using anti-His tag antibodies such as His-probe) and as purification tag.
the 6 histidine sequence has a very high affinity to nickel(II) ions and therefore is used in affinity purification columns with immobilized nickel ions. the binding of the His-Tag to the nickel ions is not efficient at acidic pH values so any purification process should be done above pH=7 (usually done at pH=8). Imidazole is used In order to elute the protein from column by competing with the His tag on binding to the immobilized Nickel ions.
His-Tag binding resins are commercially available and are widely used in recombinant protein purification processes.
this part contains the 6XHis sequence between two valine residues and terminated with the TAA stop codon.
the part can be used at the C-Terminus of the CDS with its included stop codon.
Source
synthetic construct created using the codon usage table of E.coli.
References
[http://en.wikipedia.org/wiki/Polyhistidine-tag ]