Up to what extent and how his tag effect the expression, purification and structure of a recombinant protein?
Regards
Given the hydrophilic nature of the his-tag, most of the time it will be located on the exterior of the protein structure. Sometimes the tag can be hidden, which is worthless for native protein purification.
The DNA sequence specifying a string of six histidine residues is routinely used in vectors for production of recombinant proteins. The result is expression of a recombinant protein with a 6xHis-tag fused to its N- or C-terminus.
Expressed His-tagged proteins can be purified and detected easily because the string of histidine residues binds to several types of immobilized metal ions, including nickel, cobalt and copper, under specific buffer conditions. In addition, anti-His-tag antibodies are commercially available for use in assay methods involving His-tagged proteins. In either case, the tag provides a means of specifically purifying or detecting the recombinant protein without a protein-specific antibody or probe.
To my experience there is no effect on protein expression levels, but purification is extremely simplified. Sometime the His-tag can have effects on the structure of your protein, but it is easy to insert a TEV or factor Xa cleaving site which allow you to remove the tag after purification.
In some case the His-tag can increase solubility of your protein.
His tag is not a solubility enhancer tag. It is basically used for facilitating purification of proteins by using IMAC.
Dear Thomson,
The following report gives an answer of your problems:
www.ncbi.nlm.nih.gov/pmc/articles/PMC4971304/
Best Regards
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