An expression vector tailored for large-scale, high-throughput purification of recombinant proteins.

ABSTRACT: Production of milligram quantities of numerous proteins for structural and functional studies requires an efficient purification pipeline. We found that the dual tag, his(6)-tag-maltose-binding protein (MBP), intended to facilitate purification and enhance proteins' solubility, disrupted such a pipeline, requiring additional screening and purification steps. Not all proteins rendered soluble by fusion to MBP remained soluble after its proteolytic removal, and in those cases where the protein remained soluble, standard purification protocols failed to remove completely the stoichiometric amount of his(6)-tagged MBP generated by proteolysis. Both liabilities were alleviated by construction of a vector that produces fusion proteins in which MBP, the his(6)-tag and the target protein are separ

SUBMITTER: Donnelly MI 

PROVIDER: S-EPMC2792030 | biostudies-literature | 2006 Jun

REPOSITORIES: biostudies-literature