Features of obtaining the protein complex of vegetative cultures Bacillus anthracis for proteomic mapping of strains

• Main Authors: Elena A. Koteneva, Olga I. Tsygankova, Aleksander V. Kalinin, Ivan S. Rodionov, Alena V. Abramovich, Victoriya Yu. Shcherbakova
• Format: Article
• Language: Russian
• Published: Central Research Institute for Epidemiology 2020-09-01
• Series: Журнал микробиологии, эпидемиологии и иммунобиологии
• Subjects: proteome; anthrax; bacillus anthracis; spore formation; maldi-tof/tof ms; 2d-gel electrophoresis; vegetative form; protein extraction methods
• Online Access: https://microbiol.elpub.ru/jour/article/view/870

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.