High-level expression of the Hcc domain of Clostridium botulinum neurotoxin serotype A in Escherichia coli and its immunogenicity as an antigen / 生物工程学报
Chinese Journal of Biotechnology
; (12): 812-817, 2007.
Article
em En
| WPRIM
| ID: wpr-327942
Biblioteca responsável:
WPRO
ABSTRACT
A completely synthetic gene encoding the He domain of Clostridium botulinum neurotoxin serotype A (AHc, 1287 bp, 429 aa, -50 kD) was constructed with oligonucleotides. After expressed in Escherichia coli, soluble product AHc was gained and verified by SDS-PAGE and Western blot analysis. The expressive level of recombinant AHc in E. coli was very high (36%-53% of soluble total proteins) and the purified yield was more than 30 mg/L by one-step purification. Then, the purified AHc was used to vaccinate Balb/c mice, which developed a strong and specific immune response as expected following administration of AHe protein via the subcutaneous route. Results from BoNT/A neutralization assay showed that the serum from mice vaccinated with AHc contained high titer protective antibody. These results showed that the soluble, stable and high-levelly expressive AHc not only could be produced by the prokaryotic expression system built in our lab, but also owned strong immunogenicity to prepare antitoxin for treatment and as sub-unit candidate vaccine for prophylaxis against botulinum toxin serotype A.
Texto completo:
1
Base de dados:
WPRIM
Assunto principal:
Sangue
/
Botulismo
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Proteínas Recombinantes
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Vacinas Bacterianas
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Ativação Linfocitária
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Linfócitos T
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Vacinas de DNA
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Toxinas Botulínicas Tipo A
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Clostridium botulinum tipo A
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Alergia e Imunologia
Limite:
Animals
Idioma:
En
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2007
Tipo de documento:
Article