Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2078968 | Chinese Journal of Biotechnology | 2007 | 6 Pages |
Abstract
A completely synthetic gene encoding the Hc domain of Clostridium botulinum neurotoxin serotype A (AHc, 1287 bp, 429 aa, â¼50 kD) was constructed with oligonucleotides. After being 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 AHc protein via the subcutaneous route. Results from the 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-level expressive AHc could not only be produced by the prokaryotic expression system built in the lab, but also owned strong immunogenicity to prepare an antitoxin for treatment and as a subunit candidate vaccine for prophylaxis against botulinum toxin serotype A.
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Authors
Yun-Zhou YU, Zhi-Wei SUN, Shuang WANG, Wei-Yuan YU,