Surface display of the 20-kDa N-terminal fragment of anthrax protective antigen based on attenuated recombinant <Emphasis Type="Italic">Bacillus anthracis</Emphasis> |
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Authors: | Yan-chun Wang Na Jiang De-wen Zhan Hao-xia Tao Sheng-ling Yuan Peng Wang Ling-chun Wang Zhao-shan Zhang Chun-jie Liu |
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Institution: | (1) State Key Laboratory of Pathogens and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijng, 100071, China;(2) Beijing Fisheries Research Institute, 18 Jiaomen Street, Fengtai District, Beijng, 100068, China; |
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Abstract: | Extracellular antigen 1 (EA1), a major component of the Bacillus anthracis surface layer (S-layer), was used as a fusion partner for the expression of heterologous antigen. A recombinant B. anthracis strain was constructed by integrating a translational fusion harboring the DNA fragments encoding the cell wall–targeting
domain of the S-layer protein EA1 and the 20-kDa N-terminal fragment of anthrax protective antigen (PA20) into the chromosome.
A thermosensitive plasmid expressing Cre recombinase was introduced at a permissive temperature to remove the antibiotic marker.
Cre recombinase action at the loxP sites excised the spectinomycin resistance cassette. The final derivative strains were
analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, Western blot analysis, and immunofluorescence analysis.
PA20 was successfully expressed on the S-layer of the recombinant antibiotic marker-free strain. Guinea pigs were immunized
with the attenuated recombinant B. anthracis strain, and the bacilli elicited a humoral response to PA20. This antibiotic marker-free strain and the correlative experiment
method may have potential applications for the generation of a live attenuated anthrax vaccine. |
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