title: [Gene-engineering approach for the production of A and B toxin fragments for diagnostic and immunotherapeutic use in Clostridium difficile infection].
abstract: Clostridium difficle is a causative agent of severe and difficult-to-diagnose human infections. Toxins A and B, which modify the RAS-like proteins of eukaryotic cells, are the major factor in the pathogenicity of the discussed causative agent. These very toxins are considered as the key components of the developed diagnostic and therapeutic-and-preventive preparations. The C-terminal fragments of toxins A and B as well as hybrid products, consisting of fragments of both toxins, were cloned, within the present case study, by using the pET28 plasmid vector. The recombinant plasmids were transformed into strains Escherichia Coli BL21 (DE3), and they were used later to produce the appropriate proteins. The purified protein preparations were isolated from the ultrasound bacterial-cell lysate by using the method of metal-affinity chromatography to be used later in the production of hyper-immune rabbit sera.
authors.affiliation: Gamaleya's Research Institute for Epidemiology and Microbiology, Russian Academy of Medical Sciences, Moscow, Russia.
authors.forename: N A
authors.initials: NA
authors.lastname: Varfolomeeva
authors.forename: L A
authors.initials: LA
authors.lastname: Shmarova
authors.forename: Iu F
authors.initials: IuF
authors.lastname: Belyĭ
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004761:Enterocolitis, Pseudomembranous / Q000276:immunology; D004768:Enterotoxins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D004926:Escherichia coli / Q000378:metabolism; D007106:Immune Sera; D010957:Plasmids; D015202:Protein Engineering; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; D007106:Immune Sera; D011994:Recombinant Proteins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile