A plasmid vector system for the expression of a triprotein consisting of β-galactosidase, a collagenase recognition site and a foreign gene product

S Scholtissek, F Grosse - Gene, 1988 - Elsevier
S Scholtissek, F Grosse
Gene, 1988Elsevier
A plasmid-cloning vector system has been constructed which allows the production of fusion
proteins with β-galactosidase at the N terminus, followed by a recognition sequence for the
site-specific protease, collagenase, and the foreign protein at the C terminus. A multicloning
site allows the insertion of foreign genes in any translational reading frame. Fusion proteins
were isolated by affinity chromatography on APTG-Sepharose. The foreign protein was
released from the fusion product by collagenase cleavage. The vector was successfully …
Abstract
A plasmid-cloning vector system has been constructed which allows the production of fusion proteins with β-galactosidase at the N terminus, followed by a recognition sequence for the site-specific protease, collagenase, and the foreign protein at the C terminus. A multicloning site allows the insertion of foreign genes in any translational reading frame. Fusion proteins were isolated by affinity chromatography on APTG-Sepharose. The foreign protein was released from the fusion product by collagenase cleavage. The vector was successfully utilized for the production of Escherichia coli single-stranded (ss) DNA-binding protein (SSB protein). The proteolytically released SSB protein resisted elution from an ss DNA-cellulose column with 1 M NaCl.
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