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

Gene. 1988;62(1):55-64. doi: 10.1016/0378-1119(88)90579-3.

Abstract

A plasmid-cloning vector system has been constructed which allows the production of fusion proteins with beta-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / genetics
  • Base Sequence
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics
  • Escherichia coli / genetics
  • Galactosidases / genetics*
  • Gene Expression Regulation
  • Genetic Vectors*
  • Microbial Collagenase / metabolism
  • Molecular Sequence Data
  • Plasmids*
  • Protein Processing, Post-Translational
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Proteins / genetics*
  • beta-Galactosidase / biosynthesis
  • beta-Galactosidase / genetics*

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Galactosidases
  • beta-Galactosidase
  • Microbial Collagenase

Associated data

  • GENBANK/M18897