Zinc-fingers were first known for their DNA-binding domains, consisting of multiple fingers that could bind to the major groove of DNA. Now zinc-fingers are additionally known to be a structural motif that stabilizes protein folds through the coordination of zinc atoms with cysteine and histidine residues on the protein. The zinc-finger motif is found in the myxoma virus leukemia-associated protein (MV-LAP), a protein produced by the Myxoma Virus (MV) which infects the European Rabbit. The MV-LAP protein greatly reduces immune system function in the host’s body through the down-regulation of the major histocompatibility complex class I (MHC-1) and remains undetected by the host’s immune system. Before the structure of MV-LAP can be characterized, the protein must first be expressed and purified. Two fusion proteins were cloned using two different solubility tags, Maltose Binding Protein (MBP) and NusA, as well as  a histidine affinity tag for purification. Both MV-LAP fusion proteins were successfully purified, however, when MV-LAP was cleaved from the tags, purification was unsuccessful. A third fusion protein was cloned with only the histidine tag, and no solubility tag. Without the solubility tag, however, the protein did not appear in the soluble fraction of the cell.<br/>

Expression of myxoma virus leukemia-associated protein in the presence and absence of solubility tags

253rd National Meeting (2017)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.