title: Cloning and expression in E. coli of a synthetic gene for the bacteriocidal protein caltrin/seminalplasmin.
abstract: A synthetic gene coding for the bacteriocidal protein caltrin/seminalplasmin was constructed and expressed in Escherichia coli as a fusion with beta-galactosidase. The gene was designed with a recognition site for the plasma protease, Factor Xa, coded for immediately prior to the N-terminus of caltrin. The beta-galactosidase-caltrin fusion protein was cleaved with Factor Xa to give caltrin, which was identified by its size on SDS-PAGE, its ability to react with an antiserum raised to the N-terminal nonapeptide of caltrin and its N-terminal amino acid sequence. After partial purification, synthetic caltrin was found to be active in an assay involving inhibition of growth of E.coli.
authors.affiliation: Laboratory of Molecular Biology, Medical Research Council, Cambridge, UK.
authors.forename: S
authors.initials: S
authors.lastname: Heaphy
authors.forename: M
authors.initials: M
authors.lastname: Singh
authors.forename: M J
authors.initials: MJ
authors.lastname: Gait
mesh_terms: D000970:Antineoplastic Agents; D001483:Base Sequence; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005786:Gene Expression Regulation; D005813:Genes, Synthetic; D005822:Genetic Vectors; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000138:chemical synthesis; D029584:Prostatic Secretory Proteins; D011506:Proteins / Q000235:genetics*; D029589:Seminal Plasma Proteins
chemical_list: D000970:Antineoplastic Agents; D009841:Oligonucleotides; D029584:Prostatic Secretory Proteins; D011506:Proteins; D029589:Seminal Plasma Proteins; C043076:beta-microseminoprotein