A novel family of expression vectors with multiple affinity tags for wheat germ cell-free protein expression.

ABSTRACT: BACKGROUND:Cell-free protein expression has become a widely used alternative of in vivo, cell-based systems in functional and structural studies of proteins. The wheat germ-based method outstands from the commercially available eukaryotic in vitro translation systems by its flexibility, high translation efficiency and success rate of properly folded eukaryotic protein synthesis. The original T7 promoter containing pEU3-NII vector was improved previously by addition of a ligation-independent cloning site, His6- and GST-tags, and a TEV protease cleavage site to facilitate the creation of recombinant plasmids, permit affinity purification, and enable production of purified, tag-free target proteins, respectively. RESULTS:Here, we describe a further development of pEU3-NII vector by inserting

SUBMITTER: Nagy SK 

PROVIDER: S-EPMC7071761 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature