Journal article
Production of a functional dihydrofolate synthase with a cleavable poly-His tag in Saccharomyces cerevisiae
O Patel, L Castelli, R Fernley, P Coloe, I Macreadie
Biotechnology Letters | Published : 2002
Abstract
Polyhistidine-tagged dihydrofolate synthase (DHFS) has been produced in the yeast, Saccharomyces cerevisiae, using a Cu2+-inducible expression system. The tagged DHFS is functional in vivo and was purified using immobilised metal affinity chromatography. A linker of a minimal size allows efficient cleavage of the poly-His tag using thrombin. At least 10 mg of pure DHFS can be recovered per litre of culture.