High level expression, purification and immunogenicity analysis of a protective recombinant protein against botulinum neurotoxin type E.
Botulinum neurotoxin type E heavy chain consists of two domains: N-terminal half as a translocation domain and C-terminal half (Hcc) as a binding domain. In this research a synthetic gene fragment encoding the binding domain of botulinum neurotoxin type E (BoNT/E-Hcc) was highly expressed in Escherichia coli by pGEX4T-1 vector. After purification, the recombinant BoNT/E-Hcc was evaluated by SDS-PAGE and western blot (immunoblot) analysis. Average yields obtained in this research were 3.7 mg recombinant BoNT/E-Hcc per liter of bacterial culture. The recombinant protein was injected in mice for study of its protection ability against botulinum neurotoxin type E challenges. The challenge studies showed that, vaccinated mice were fully protected against 10⁴ × minimum lethal dose of botulinum neurotoxin type E.
Biotechnology Department, Institute of Basic and Applied Sciences, Cukuruva University, Adana, Turkey, va.cell@yahoo.com.
Ebrahim
E
Valipour
Mir-Latif
ML
Moosavi
Jafar
J
Amani
Shahram
S
Nazarian
D000818:Animals; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D001905:Botulinum Toxins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D001906:Botulism / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D003014:Clostridium botulinum / Q000235:genetics / Q000276:immunology / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D001428:Bacterial Vaccines; D011994:Recombinant Proteins; D001905:Botulinum Toxins; C096326:botulinum toxin type E