Evaluation of 23 kDa far C-terminal portion of Clostridium botulinum toxin type E as a candidate for recombinant vaccine : 클로스트리디움 보툴리눔 E 형 독소의 23 kDa C-말단에 대한 재조합 백신 후보물질로서의 평가
The Graduate School of Sunmoon University
Department of Biological Sciences
recombinant vaccine 23 kDa far C-terminal botulinum Clostridium toxin type E;
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Botulinum neurotoxin (BoNT) is a microbial product synthesized by the organisms Clostridium botulinum, Clostridium baratii, and Clostridium butyricum. C. botulinum produces all seven known serotypes (A to G), whereas C. baratii and C. butyricum produce only one serotype each (F and E, respectively). Clostridium botulinum neurotoxin type E (BoNT/E) is produced by C. botulinum serotype E. BoNTs are comprised of a 150 kDa dichain protein with a 50 kDa light chain linking to a 100 kDa heavy chain through a disulfide bridge. Heavy chain enable the toxin to bind to neuronal cell by action of 50 kDa C-terminal and to make translocation channel by the action of 50 kDa N-terminal, whereas the light chain accounts for catalytic activity. In this work, 23 kDa far C-terminal portion of heavy chain, which is designated as EHc23, was cloned into pGEX-4T-1 expression vector, expressed in E.coli BL21-RIL and purified by using GST-affinity column. Mice were immunized with 30 μg of GST-free EHc23 protein three times repeatedly, at 15 day intervals. In vivo BoNT/E challenge test showed that the immunization prolonged mouse survival against BoNT/E treatment.