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Simplifying protein expression with ligation-free, traceless and tag-switching plasmids.

Protein Expr Purif.. 2012-09;  85(1):9-17
Durani V, Sullivan BJ, Magliery TJ. a Department of Chemistry, The Ohio State University, Columbus, OH 43210, USAb Ohio State Biochemistry Program, The Ohio State University, Columbus, OH 43210, USAc Department of Biochemistry, The Ohio State University, Columbus, OH 43210, USA
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摘要

Synthetic biology and genome-scale protein work both require rapid and efficient cloning, expression and purification. Tools for co-expression of multiple proteins and production of fusion proteins with purification and solubility tags are often desirable. Here we present a survey of plasmid vectors that provide for some of these features with a focus on tools for rapid cloning and traceless tagging – a setup that facilitates removal of fusion tags post-purification leaving behind no ‘scar’ on the final construct. Key features are reviewed, including plasmid replication origins and resistance markers, transcriptional promoters, cloning methods, and fusion tags and their removal by proteolysis.... More

关键词

Plasmid; Protein expression; Co-expression; Protease cleavage; Ligation independent cloning; Traceless tagging