title: Production and cell surface display of recombinant anthrax protective antigen on the surface layer of attenuated Bacillus anthracis.
abstract: To investigate the surface display of the anthrax protective antigen (PA) on attenuated Bacillus anthracis, a recombinant B. anthracis strain, named AP429 was constructed by integrating into the chromosome a translational fusion harboring the DNA fragments encoding the cell wall-targeting domain of the S-layer protein EA1 and the anthrax PA. Crerecombinase action at the loxP sites excised the antibiotic marker. Western blot analysis, fluorescence-activated cell sorting and immunofluorescence analysis confirmed that PA was successfully expressed on the S-layer of the recombinant antibiotic marker-free strain. Notwithstanding extensive proteolytic degradation of the hybrid protein SLHs-PA, quantitative ELISA revealed that approximately 8.1 × 10(6) molecules of SLHs-PA were gained from each Bacillus cell. Moreover, electron microscopy assay indicated that the typical S-layer structures could be clearly observed from the recombinant strain micrographs.
authors.affiliation: State Key Laboratory of Pathogens and Biosecurity, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai District, Beijing, 100071, China, springwyc@gmail.com.
authors.forename: Yan-chun
authors.initials: YC
authors.lastname: Wang
authors.forename: Sheng-ling
authors.initials: SL
authors.lastname: Yuan
authors.forename: Hao-xia
authors.initials: HX
authors.lastname: Tao
authors.forename: Ling-chun
authors.initials: LC
authors.lastname: Wang
authors.forename: Zhao-shan
authors.initials: ZS
authors.lastname: Zhang
authors.forename: Chun-jie
authors.initials: CJ
authors.lastname: Liu
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D001408:Bacillus anthracis / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014613:Vaccines, Attenuated / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; C055764:S-layer proteins; D014613:Vaccines, Attenuated; C030325:anthrax toxin