Affiliation:
1. Stavropol Research Antiplague Institute
Abstract
Introduction. The study of the protein composition of the causative agent of anthrax — Bacillus anthracis, allows you to identify both general species and individual characteristics of strains that differ in phenotypic properties, manifested mainly in the vegetative form and which are important for virulence, immunogenicity and the ability to adapt to different vegetation conditions.Purpose of the work. In the group of anthrax microbe strains having different plasmid composition and virulence, different methods of extraction of the total proteome from vegetative bacillus cells have been tested.Results. In the course of the work, it was shown that the rate of spore formation varies significantly between individual strains of the anthrax microbe and can have a significant impact on the efficiency of extraction and the composition of the protein complex. Preliminary treatment with lysozyme, which affects the cell membrane, promotes a more complete lysis of cells, and ultramicrocentrifuge filtration provides complete specific sterility of the obtained samples.Conclusion. A culture preparation scheme was developed for B. anthracis, which allows one to obtain a culture in the vegetative phase of the life cycle and to efficiently extract proteins in combination with reliable disinfection of samples.
Publisher
Central Research Institute for Epidemiology
Reference6 articles.
1. Castanha E.R., Fox A., Fox K.F. Rapid discrimination of Bacillus anthracis from other members of the B. cereus group by mass and sequence of «intact» small acid soluble proteins (SASPs) using mass spectrometry. J. Microbiol. Methods. 2006; 67(2): 230-40. DOI: http://doi.org/10.1016/j.mimet.2006.03.024
2. Lasch P., Beyer W., Nattermann H., Stämmler M., Siegbrecht E., Grunow R., et al. Identification of Bacillus anthracis by using matrix-assisted laser desorption ionization — time of flight mass spectrometry and artificial neural networks. Appl. Environ. Microbiol. 2009; 75(22): 7229-42. DOI: http://doi.org/10.1128/AEM.00857-09
3. Mottaz-Brewer H., Norbeck A., Adkins J., Manes N., Ansong C., Shi L., et al. Optimization of proteomic sample preparation procedures for comprehensive protein characterization of pathogenic systems. J. Biomol. Tech. 2008; 19(5): 285-95.
4. Chitlaru T., Shafferman A. Proteomic studies of Bacillus anthracis. Future Microbiol. 2009; 4(8): 983-98. DOI: http://doi.org/10.2217/fmb.09.73
5. Gao Z., Wang Z., Zhang K., Li Y., Zhang T., Wang D., et al. Experimental validation of Bacillus anthracis A16R proteogenomics. Sci. Rep. 2015; 5: 14608. DOI: http://doi.org/10.1038/srep14608