title: [Effective expression of fragments of a botulinum neurotoxin type A gene, coding for the L-chain and H-chain in E. coli, with formation of products causing protective immunity to administration of the toxin].
abstract: Native Clostridium botulinum gene coding for type A neurotoxin has been used to construct recombinant derivatives coding separately for L and H polypeptide chains of the toxin. The gene derivatives have been cloned into an expression vector pET28b in E. coli BL21 (DE3) cells. The recombinant L and H proteins seem to be the major individual proteins after IPTG induction of the recombinant cells. Each of the proteins has been accumulated only in inclusion bodies. The recombinant L chain (but not H chain) has been successfully resolubilized. Each of the proteins contains six His residues on the N terminus which allows purification on Ni-agarose columns with high yield. No toxic effect has been observed for both L and H chains after injection of 10 micrograms of recombinant preparations purified from inclusion bodies. Moreover, the injection resulted in an increase in the titer of specific antibodies which protected mice from 1 DLM of type A native botulinum neurotoxin. Hence, the recombinant neurotoxin protein derivatives which are present in E. coli inclusion bodies can be a source of material for producing diagnostic and therapeutic sera against type A botulinum neurotoxin.
authors.affiliation: N. F. Gamaleya Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences, Moscow, Russia.
authors.forename: Iu V
authors.initials: IuV
authors.lastname: Vertiev
authors.forename: A G
authors.initials: AG
authors.lastname: Zdanovskiĭ
authors.forename: S A
authors.initials: SA
authors.lastname: Borinskaia
authors.forename: T
authors.initials: T
authors.lastname: Martin
authors.forename: E L
authors.initials: EL
authors.lastname: Gening
authors.forename: N K
authors.initials: NK
authors.lastname: Iankovskiĭ
mesh_terms: D001483:Base Sequence; D001905:Botulinum Toxins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D004269:DNA, Bacterial; D011994:Recombinant Proteins; D001905:Botulinum Toxins