[Properties of pectolitic phytopathogenic bacteria isolates obtained in Ukraine].

Bacteria obtained from potato tubers having symptoms of soft rot and grown in different regions of Ukraine are identified as Pectobacterium carotovorum subsp. carotovorum. These bacteria strains are able to produce bacteriocines. Their killer activity in respect of P. carotovorum and Esherichia coli has been studied. The sensitivity to bactericines has been shown. Purified fractions of bacteriocines having high molecular weight (MCTV) have been obtained. The difference in composition of proteins from phage tails as compared to the ones in P. carotovorum J2 has been studied by the method of electrophoresis. It was found that the composition of MCTV major proteins of studied isolates mostly corresponds to P. carotovorum J2. The set of enzyme minor fractions has some different compositions as compared to P. carotovorum J2. It has been hypothesized that this difference is responsible for killer specificity.

[Matrix activity of mRNP of different cell localization from plants infected by minus-genome curly potato dwarfness virus in protein-synthesizing system in vitro].

Using structures of mRNP of different cell localization isolated from plants infected by minus-genomecurly potato dwarfness virus (CPDV), it was established that synthesis of virus proteins in vitro is mainly realized on membrane-related and free polysomal mRNP capable to direct synthesis of proteins programmed in RNA in cell-free protein-synthesizing systems. The obtained results prove that synthesis of virus proteins is actively realized on membrane-related nonpolysomal mRNP - 23520 imp/min. An analogous distribution of matrix activity is observed also in the case when mRNA isolated from mRNP of different cell localization was introduced in protein-synthesizing system. The least matrix activity was observed in cytoplasmic nonpolysomal mRNPs which are low-active in the translation system in vitro - only 1890 imp/min. The function of free cytoplasmic nonpolysomal mRNP is apparently mainly reduced to the transport and reserve of genetic information in a cell.

[Effect of structural P-protein of potato curly dwarf virus on replicase activity of mRNP-complexes of infected plants].

All representatives of rhabdoviruses contain a nucleocapside phosphoprotein - P-protein which is an essential subunit of the viral RNA-dependent RNA polymerase complex. As a result of studying the effect of nucleocapside protein P(NS) on replicase activity of mRNP isolated from plants infected by potato curly dwarf virus in the system in vitro, it was established that nucleocapside P-protein stimulates considerably the replicase activity of membrane-bound polysomal m-RNP P-protein being available in concentration of 15 microg/ml in the replication system in vitro of membrane-bound polysomal mRNP, the replicase activity increased 11.7 times. This property of nucleocapside P-protein at the same concentration was displayed to a less extent with the presence of free polysomal mRNP, in the system in vitro. Thus the replicase activity mRNP-complexes in the replication system in vitro depends on the presence of nucleocapside viral P-protein in the system. Its concentration being increased or decreased, one can observe the change of the replicase activity.

[Functional properties of prosomal protein 39 kappaDa from Datura stramonium leaves infected with Potato Virus X].

Free cytoplasmic informosomes isolated from Datura stramonium plants infected by PVX contain a low-molecular ribonucleoprotein complex (RNP). This complex as to its main physico-chemical parameters (sedimentation coefficient 10S, buoyant density in CsSO4 1.31 g/cm3, stability to 1% lauroylsarcosinate-Na) corresponds to the prosome (inhibitory RNP). Prosomes isolated from free mRNP of D. stramonium plants infected by PVX contain the protein of 39 kDa. This protein was shown to be capable to phosphorylate in vitro in the composition of informosomes and prosomes. It is possible that this protein can be the protein-repressor, since it is absent in the translated polysome-associated form of mRNP. The label incorporation has shown that the protein of 39 kDa is able to reduce in vitro the template activity of genomic RNA PVX to 40% and RNA TMIV--to 30%. Moreover, the protein 39 of kDa has the protease activity. It affects substrate-case in like trypsin. It is supposed that it can participate in splitting the intracellular proteins as well as in the expression of the virus genome, it can also influence the template activity of cell RNAs.

[Antigenic relationship between nucleocapsid proteins of phyto- and zoorhabdoviruses].

The methods of electrophoresis in PAAG and immunological method were used for comparative analysis of structural proteins of phytorhabdovirus of potato curly dwarf (PCDV) and zoorhabdoviruses-vesicular stomatitis virus (VSV) and fixed rabies Virus (RV). Molecular weight of viral proteins was determined by the method of polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The proteins with molecular weight 45-51 kD, are probably, the major component of the viral nucleocapsid. Nucleocapsid protein 45 kD RV virus was isolated by the method of preparative electrophoresis and then the monospecific serum was obtained. The Ouchterlony and immunoblotting method were used to show, that nucleocapsid proteins with molecular weights 51 and 45 kD both of phytorhabdovirus PCDV and zoorhabdoviruses VSV and RV are serologically related. The obtained data may be used in biotechnology as the basis for creation of a new class of diagnostic preparations with the purpose to detect RV virus using proteins of curly potato dwarf virus and may be also used in serological tests to reveal viruses of Rhabdoviridae family in various eukaryotic objects.

[Immunospecific detection of the potato virus X by the surface plasmon resonance].

Highly specific and proximal method of laboratory diagnostics of the viral disease, has been developed using the structural protein of the potato virus X, as a model, and monospecific antibodies to it. The immunospecific determination of the potato virus X was carried out by the surface plasmon resonance method using specific IgG-antigen complexes, immobilized on the sensor surface modified by rodanide and protein A of Staphylococcus aureus.

[Enzyme activity of nucleocapsid proteins of the curly potato dwarf virus]. / Fermentativnaia aktivnost' nukleokapsidnykh belkov fitorabdovirusa kurchavoi karlikovosti kartofelia.

The individual nucleocapsid proteins of phytorhabdovirus of curly potato dwarf were isolated: N-protein (m. w. 56 kDa), NS-protein (m. w. 49 kDa), L-protein (m. w. 128 kDa). It was establish that L and NS proteins displayed enzyme activity in the replication, phosphorylation and adenylation systems in vitro. Major N protein (m. w. 56 kDa) did not show the enzyme activity in these systems. The revelation of RNA-polymerase, poly(A)-polymerase and proteinkinase activities of nucleocapsid proteins are characteristic of the viruses belonging to the Rhabdoviridae family.

[Physico-chemical properties and functional peculiarities of mRNP of different subcellular location of Nicotiana rustica plants affected by potato curly dwarf virus]. / Fiziko-khimicheskie svoistva i funktsional'nye osobennosti mRNP razlichnoi subkletochnoi lokalizatsii rastenii Nicotiana rustica, porazhennykh virusom kurchavoi karlikovosti kartofelia.

mRNP of different subcellular location of N. rustica plants affected by PCDV contain virus-specific proteins. RNA-replicase, poly (A)-polymerase and protein kinase have been found in mRNP under study. The most expressed replicase, poly(A)-polymerase and protein kinase activity has been noticed in the systems containing membrane-bound non-polysomal mRNP.