Small-scale, semi-automated purification of eukaryotic proteins for structure determination.

ABSTRACT: A simple approach that allows cost-effective automated purification of recombinant proteins in levels sufficient for functional characterization or structural studies is described. Studies with four human stem cell proteins, an engineered version of green fluorescent protein, and other proteins are included. The method combines an expression vector (pVP62K) that provides in vivo cleavage of an initial fusion protein, a factorial designed auto-induction medium that improves the performance of small-scale production, and rapid, automated metal affinity purification of His8-tagged proteins. For initial small-scale production screening, single colony transformants were grown overnight in 0.4 ml of auto-induction medium, produced proteins were purified using the Promega Maxwell 16, and purifica

SUBMITTER: Frederick RO 

PROVIDER: S-EPMC2668602 | biostudies-literature | 2007 Dec

REPOSITORIES: biostudies-literature