Abstract
We developed epitope-tagged metallothionein IIA cDNA by the addition of an 8-amino acid sequence, DYKDDDDK, at the N-terminus metallothionein using polymerase chain reaction. The fusion proteins were expressed by transfection in human embryonic kidney 293 (HEK-293) cells and were specifically recognized by commercially available antibody, an anti-Flag M2 monoclonal antibody. Furthermore, the fusion protein was detectable on a Western blot, whose migration was expectedly at 7000-8000 in an SDS-denatured form. Thus, availability of the epitope-tagged metallothionein will facilitate the investigation of cellular mechanisms of the protein such as its nuclear localization mechanism.
