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: Yan-chun Wang, Sheng-ling Yuan, Hao-xia Tao, Ling-chun Wang, Zhao-shan Zhang, Chun-jie Liu
• Journal: World journal of microbiology & biotechnology (World J Microbiol Biotechnol) Vol. 31 Issue 2 Pg. 345-52 (Feb 2015) ISSN: 1573-0972 [Electronic] Germany
• PMID: 25504373 (Publication Type: Journal Article)

Chemical References

• Antigens, Bacterial
• Bacterial Toxins
• Membrane Glycoproteins
• Recombinant Fusion Proteins
• S-layer proteins
• Vaccines, Attenuated
• anthrax toxin

Topics

• Antigens, Bacterial (genetics, immunology, metabolism)
• Bacillus anthracis (genetics, immunology, metabolism)
• Bacterial Toxins (genetics, immunology, metabolism)
• Cell Membrane (metabolism)
• Cloning, Molecular
• Membrane Glycoproteins (genetics, metabolism)
• Mutation
• Recombinant Fusion Proteins (genetics, immunology, metabolism)
• Sequence Analysis, DNA
• Vaccines, Attenuated (immunology)