Publications on the project |
112 Особенности морфогенетического развития вирулентных мутантов єрвиниофага ZF40 |
Authors: | Korol N.A., Romaniuk L.V., Ostapchuk A.M., Ivanytsya T.V., Kushkina A.I, Tovkach F.I. | |
Summary: | | |
Keywords: | | |
Edition: | | | | 2011,
,Russian |
112 Polypeptide content and HPLC-chromatography of the virions of phage ZF40 mutants |
Authors: | Ostapchuk A.M., Korol N.A., Romaniuk L.V., Tovkach F.I. | |
Summary: | Study of the polypeptide content of erwiniophage ZF40c5/5 and ZF40-421 virulent mutants has shown that their virions include no less than 10 structural proteins with molecular weights ranging from 16.9 to 96.5 kDa. Three polypeptides belong to a group of major proteins with molecular weights 39.2, 33.1 and 18.5 kDa. They correlate with the polypeptides of phage head, tail sheath and tail core correspondingly. It has been proven that the protein contents of these phages are identical, taking into account that the percent ratio of all polypeptides approaches 1.0. The polypeptide profile of isogenic variant of phage ZF40-421 obtained on EccRC5297 is
characterized by another ratio of major proteins. These differences are reflected in the structure of procapsids, that explains low level of stability and viability of the variant.
The work shows for the first time the possibility of using HPLC-chromatography for studying native phage particles and their structural components. | |
Keywords: | Erwinia carotovora, bacteriophage ZF40, phage mutants, virion polypeptides, HPLC-chromatography. | |
Edition: | | | | 2011,
,Russian |
112 CHROMATOGRAPHIC PROFILES AND ELECTRON MICROSCOPY OF VIRUS-LIKE PARTICLES (VLP) OF ERWINIA CAROTOVORA SUBSP. CAROTOVORA |
Authors: | Ivanytsya T.V., Krylova K.D., Sergeeva Zh.U., Tovkach F.I. | |
Summary: | Incomplete or defective bacteriophages of Erwinia carotovora subsp. carotovora (Ecc) are a collection of individual virus-like particles (VLP), which are formed of a virion: phage tails, capsid formations and the basal plates. In addition, Ecc-induced lysates may contain other types of VLP: individual cores and sheaths, polycores, and polysheaths. To analyze the entire set of particle chromatographic analysis on DEAE cellulose fiber 23 SS was proposed and was followed by electron microscopy of VLP. It was established, that the chromatographic profiles of VLP which are able to kill sensitive bacteria, have unique and conserved features of polylysogenic strains of E. carotovora. However, in the case of isogenic strains they are completely identical. While using a particular set of indicator strains, VLP-profiles can characterize not only defective, but also the true lysogeny of erwinia. Electron microscopy studies showed that at the same VLP-profiles of two isogenic strains, the number and types of phage particles may differ significantly. In the Ecc-induced lysate 48A-7/4b not only an excess of phage basal plates was found, but also a new type of phage tails. In this strain there are no other changes except for Tn9 transposition into a cryptic plasmid pCA25, therefore it may be assummed that this small extrachromosome affects significantly, the replication of bacterial DNA, and consequently the overall dynamics of the E.carotovora chromosome. | |
Keywords: | Erwinia carotovora, defective polylysogeny, virus-like particles, chromatography, and electron microscopy. | |
Edition: | | | | 2011,
,Russian |
112 Long-term preservation of unstable bacteriophages of Enterobacteria |
Authors: | Tovkach F.I., Zhuminska G.I., Kushkina A.I. | |
Summary: | Bacteriophages are integral components of bacterial communities. Their practical applications and significance for humans are various. Thus, keeping phage collections along with their specifi c host bacteria is an urgent and important mission for biologists. The problems of the long-term storage of phages steel are not completely solved. The main diffi culties may occur due to the structural instability of virions as well as an accelerated genetic variability in phage genomes both in vivo and in vitro. In the paper the results of 10-years observation over unstable bacteriophage storage process was presented as well the method of their long-term preservation was proposed. It consisted of the optimization of STMG buffer system (200 mM NaCl, 10 mM Tris-HCl, pH 7.4, 1 mM MgCl2, 100 μg/ml gelatin) [Serwer P., Pichler M.E. Electrophoresis of bacteroiphage T7 capsids in agarose gels// J.Virol. – 1978. – V.28, N3. – P.917-928] by higher gelatin concentration or its replacement by fi coll (2 – 6 %), and increasing of Mg2+ concentration to 10 mM. The proposed buffer composition allowed saving structural entirety of unstable phage virions during their long-term storage. | |
Keywords: | Bacteriophages, long-term storage, structural instability, gelatine,fi coll. | |
Edition: | | | | 2012,
,English |
112 New Approach for Identification of Bacteriophage Virion Structural Proteins |
Authors: | Korol N.A., Tovkach F.I. | |
Summary: | The ability of the phage structural polypeptides to undergo post-translational modification makes the task of correlation of the primary nucleotide sequence data with the actual structural proteins of a virion extremely challenging. This study describes an alternative model approach based on two-stage chromatography for allocation of virion structural components and identification of their major polypeptides. Bacteriophage T4D, its amber mutant T4D23 (amH11) and its tail preparations were purified, concentrated and separated by ion exchange chromatograpgy based on fibrous DEAE-cellulose. The major tail fraction was then exposed to size-exclusion chromatography which enabled to separate tail components by size. This method proved itself as a highly efficient and gentle enough to save most of the biological material without changing the basic properties of the native phage. The result also shows that the accumulation of individual phage tails in the course of the amber mutant T4D23 (amH11) propagation on the permissive host Escherichia coli CR63 was resulted by changes in the conditions of reproduction. The ability of bacteriophages to form an excess of tails, capsids and other structures during reproduction on a non-traditional host provides an alternative way for obtaining highly concentrated preparations of virion components for further analysis of their major proteins and determination of the genes responsible for their synthesis. | |
Keywords: | Bacteriophage T4, amber mutant, virion components, ion-exchange, size-exclusion chromatography, polypeptides.
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Edition: | | | | 2013,
,English |
The events in the framework of the project |
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112 4.2. Physical and chemical basis of synthesis and formation properties of nanomaterials medical Purpose: Expected results:Release of new product: material Stage 1:Здійснення кількісної оцінки природних фагових популяцій на наявність в їх складі аберантних біологічних наноструктур типу прокапсидів та мініпрокапсидів, які не містять молекул ДНК і РНК Stage 2:Одержання та аналіз фагових мутантів, які при абортивних інфекціях синтезують максимальну кількість прокапсидних наноструктур Stage 3:Стандартизація умов одержання, очистки і зберігання фагових прокапсидів Stage 4:Біофізичні, біохімічні і молекулярно-біологічні параметри фагових прокапсидів Stage 5:Визначення впливу ікосаедричних фагових прокапсидних наноструктур на імунну систему макроорганізму зокрема на В- і Т-лімоцити експериментальних тварин
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