Scientific Journal of KubSAU

Polythematic online scientific journal
of Kuban State Agrarian University
ISSN 1990-4665
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Sakhabutdinova Lyalya Renatovna

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Institute of Biochemistry and Physiology of Microorganisms

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abstract 1341710034 issue 134 pp. 404 – 426 29.12.2017 ru 1381
A method of obtaining insertion mutants for the hoc gene, which encodes for the main phage antigen, was developed on the model of bacteriophage T4. This gene was cloned in the plasmid pBSL0+ and was disrupted by insertion of foreign DNA. The phage mutants were obtained by in vivo phage-plasmid recombination. The construction of insertion bacteriophage mutants was carried out in two stages. The resulting mutants on this procedure could be grown on wild-type E. coli strains, which is convenient for the production and use of these phages in therapy. The mutants obtained had reduced antigenicity. At the same time, the yield of the mutant strains was high when they were grown on the non-suppressor E. coli laboratory strains. A number of stages of purification of the bacteriophage mutants obtained were performed. Preparations have been studied by transmission electron microscopy and mass spectrometry. By several periodic cultivations of the mutant bacteriophages, it was shown that mutations of this type are stably maintained during more than 50 generations. T4 related bacteriophages of the family Myoviridae, for example, T-even, have the significant homology amongst their genomes, which makes possible to produce similar mutants. Thus, our method was developed to obtain mutants with reduced antigenicity which can be used for both the treatment of systemic infections, and diarrhea in the case, when, bacteriophages penetrate into the bloodstream. Such phages can be used in medicine and veterinary. The reported study was partially supported by RFBR, research projects No. 13-04-00991, 16-44-230855