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Surface display of the 20-kDa N-terminal fragment of anthrax protective antigen based on attenuated recombinant Bacillus anthracis

文献类型： 外文期刊

作者： Wang, Yan-chun 1 ; Jiang, Na 1 ; Zhan, De-wen 1 ; Tao, Hao-xia 1 ; Yuan, Sheng-ling 1 ; Wang, Peng 1 ; Wang, Ling-chun 1 ;

作者机构： 1.Beijing Inst Biotechnol, State Key Lab Pathogens & Biosecur, Beijing 100071, Peoples R China

2.Beijing Fisheries Res Inst, Beijing 100068, Peoples R China

关键词： DNA;loxP site;Cre recombinase: expression;spectinomycin: 1695-77-8;anthrax protective antigen: PA20;expression;N-terminal fragment;extracellular antigen 1: EA1;surface layer protein;cell wall-targeting domain;thermosensitive plasmid: antibiotic marker;humoral response;antigen surface display;drug resistance casette

期刊名称：WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY （ 影响因子：3.312； 五年影响因子：3.58 ）

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收录情况： SCI

摘要： 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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