title: Reconstitution of active recombinant Shiga toxin (Stx)1 from recombinant Stx1-A and Stx1-B subunits independently produced by E. coli clones.
abstract: Escherichia coli clones expressing recombinant Shiga toxin (Stx)1-A and recombinant Stx1-B subunits, were established. Culture supernatants of these clones were examined for inhibitory activity on in vitro protein synthesis using luciferase as a reporter enzyme. Culture supernatant of the clone expressing Stx1-A, but not Stx1-B, showed the inhibitory activity. Neither recombinant Stx1-A nor Stx1-B showed Vero cell cytotoxicity. For reconstitution of biologically active toxin, the culture supernatants of the Stx1-A clone and the Stx1-B clone were mixed. The reconstituted recombinant Stx1 showed both Vero cell cytotoxicity and inhibition of in vitro protein synthesis.
authors.affiliation: Shionogi Research Laboratories, Shionogi and Co. Ltd., Osaka, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nishikawa
authors.forename: J
authors.initials: J
authors.lastname: Fujii
authors.forename: S
authors.initials: S
authors.lastname: Yoshida
authors.forename: T
authors.initials: T
authors.lastname: Yutsudo
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D001427:Bacterial Toxins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods; D003603:Cytotoxins / Q000032:analysis / Q000378:metabolism; D003956:Dialysis; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D017353:Gene Deletion; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D022603:Shiga Toxins; D014709:Vero Cells
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D003603:Cytotoxins; D011994:Recombinant Proteins; D022603:Shiga Toxins