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
in English
| WPRIM
| ID: wpr-327942
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Blood
/
Botulism
/
Recombinant Proteins
/
Bacterial Vaccines
/
Lymphocyte Activation
/
T-Lymphocytes
/
Vaccines, DNA
/
Botulinum Toxins, Type A
/
Clostridium botulinum type A
/
Allergy and Immunology
Limits:
Animals
Language:
English
Journal:
Chinese Journal of Biotechnology
Year:
2007
Type:
Article
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