[pSTS4--a new shuttle vector for cyanobacterium Plectonema boryanum CALU 465]. / pSTS4--novyi chelnochnyi vektor dlia tsianobakterii Plectonema boryanum CALU 465.

Three vector constructions--pSTS2, pSTS3 and pSTS4 have been obtained on the basis of the standard vector for Escherichia coli pBR 322, cyanobacterial plasmid pSM1 and its separate fragments. The structure of pSTS4, carrying the full-size plasmid pSM1 (cloned by the restriction site Hpa II), replicated both in the cells of E. coli and in the cells-transconjugants of the trichomal cyanobacterium Plectonema boryanum. Under these conditions resistance of P. boryanum cells to ampicillin increases three orders, and to tetracyclin--50 times. Structural integrity of pSTS4 is stably supported in conjugates during seven successive passages (time of the experiment) and preserved the transforming capacity in respect of the cells of E. coli. It is supposed that pSTS4 is a shuttle-vector with two active origins of replication; one of them provides for the replication of pSTS4 in E. coli cells, another--in P. boryanum cells.

[The nucleotide sequence of the rep gene of cyanobacterial plasmid pSM1]. / Nukleotidnaia posledovatel'nost' gena Rep-belka tsianobakterial'noi plazmidy pSM1.

The nucleotide sequence was established for the rep gene of plasmid pSM1 isolated from cyanobacterium Plectonema boryanum CALU 465. Both nucleotide sequence and the encoded amino acid sequences showed 98% homology to the corresponding sequences of small plasmids pPF1, pGL3, pPBS1, pBLX, and pPB1. An active center was identified in the replicative protein sequences.

[Isolation and purification of nonspecific nuclease of cyanobacterium Plectonema boryanum CALU 465]. / Vydelenie i ochistka nespetsificheskoi nukleazy tsianobakterii Plectonema boryanum CALU 465.

Nonspecific nuclease has been isolated from the cells of cyanobacterium Plectonema boryanum and purified to homogenic state. It has been established that the method of centrifugation of cell-free culture extract in the sucrose density gradient is efficient for the separation of pigment proteins and enzyme concentration. Under the successive use of two ion-exchangers the nuclease activity was determined in the concentration range of NaCl 0.065-0.085 M after separation of the cell-free cyanobacterium extract on the column with phosphocellulose in the range of 0.2-0.25 M, on the column with DEAE--Toyopearl respectively. The molecular mass of nuclease which is 40 kDa, has been determined by electrophoresis in polyacrylamide gel under denaturating conditions and gel-filtration on Sephadex G-100. It has been also established that the given enzyme is monosubunitary as to its structure.

[Phagoresistance of filamentous cyanobacteria clones]. / Priroda fagorezistentnosti klonov nitchatykh tsianobacterii.

The paper deals with formation regularities of phagoresistant clones of cyanobacteria in two productive virus-cell systems: heterocyst cyanobacterium Nostoc linckia--cyanophage N-2, and mutant in heterocysts strain of Anabaena variabilis--cyanophage A-1. Frequency of spontaneous formation of phagoresistant clones of cyanobacterium N. linckia varies within 1.0-8.0 x 10(-6) per a cell, A. variabilis--5.0 x 10(-6)-7.0 x 10(-7) per cell. All the studied phagoresistant clones of N. linckia have identical morpho-cultural properties and do not differ from those of the initial culture. Phagoresistant clones of A. variabilis are presented by two groups. One of them, as to its properties, does not practically differ from the wild type culture. The second group differs considerably from the initial culture A. variabilis as to a number of characteristics--time of colonies appearance, their amount, length of trichomas, specific rate of growth and biomass accumulation. Spontaneous transfer of cyanophages to the culture liquid of clones resistant forms of cyanobacteria has not been revealed. Lysis of cells of the studied clones also was not induced under the effect of mytomycin C, thermal treatment and UV-irradiation. Cyanophage N-2 is not adsorbed by the cells of resistant cloned forms of cyanobacteria N. linckia. Only nonspecific adsorption takes place on the cells of phage-resistant clones of A. variabilis of both groups: about 20% of virions introduced in the adsorption mixture. Basing on the data obtained, it is supposed that phage-resistance of stable clones of filamentous cyanobacteria under the conditions of the given experiment is determined by the structure modification of cells receptors.

[Construction of vector plasmids for the Plectonema boryanum cyanobacterium and creation, on such basis, of the vector-host system]. / Konstruirovanie vektornykh plazmid dlia tsianobakterii Plectonema boryanum i sozdanie na ikh osnove sistemy vektor-khoziain.

The plasmid composition of the Plectonema boryanum Gom. Cyanobacterium, strain CALU 465, was analyzed. A small pSM1 plasmid, size 1.5, was chosen for genetic engineering. The physical map, constructed for the plasmid, was used to create the pSTS series, i.e. vector construction for the P. boryanum cyanobacterium. A method was selected and tested for the introduction of the vector into host cells. The needed orientation of pSM1 cyanobacterium plasmid within the vector as well as a possible role of its separate elements in inheriting the construction by host were defined.

[Comparative characteristics of native proteinases of the cyanobacteria Plectonema boryanum and Anabaena variabilis and those induced by cyanophages]. / Porivnial'na kharakterystyka natyvnykh ta indukovanykh tsianofahamy proteïnaz tsianobakterii Plectonema boryanum i Anabaena variabilis.

Physico-chemical and catalytic properties of proteinases of native and induced cells of cyanobacteria Plectonema boryanum have been comparatively studied. It has been established that at early stages of reproduction of cyanophage LPP-3 in cyanobacteria P. boryanum is formed de novo proteinase complex consisting at least of five enzymes. Proteinases induced by the virus are distinguished from those of native cells by a series of physico-chemical characteristics and possess higher catalytic activity. Analogous virus-induced changes in proteinase complex also occur in the system cyanobacterium Anabaena variabilis--cyanophage A-1. Possible functions of certain enzymes of proteinase complex in the virus pathology of cyanobacteria cells are discussed in the paper.

[Development of cyanobacterial phages at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine (History and perspectives)]. / Razvitie tsianofagii v Institute mikrobiologii i virusologii NAN Ukrainy (Istoriia i perspektivy).

The paper deals with the basic trends of fundamental investigations of the Department of Algae Viruses in the field of cyanophagia-ecology, biological and physico-chemical properties of cyanophages as well as interrelation with the host cells. Such problems as a possibility to use the system cyanophage-cyanobacteria as the experimental model for development of the unified functional model of productive infection, efficient methods of prophylaxis and therapy of virus infections as well as the solution of various biotechnological problems are discussed.

[Localization of DNAase activity in cyanobacterium Plectonema boryanum CALU 465]. / Lokalizatsiia DNKaznoi aktivnosti v kletkakh tsianobakterii Plectonema boryanum CALU 465.

It was shown in experiments carried out on cyanobacterium P. boryanum, that when cells are incubated in a buffer with low concentrations of nonion detergent (0.1%, Triton X-100) one can observe the DNAse activity release into the medium. The methods of supersound desintegration of cells and following gradient centrifugation of the cell-free extracts were used to obtain three fractions of the cell membranes identified by means of spectrophotometric analysis. It has been established that P. boryanum is characterised by high level of DNAse activity associated with the cytoplasmic cell membrane. As to its activity character the DNAse under study belongs to the class of nonspecific endonucleases.

[Physical mapping of DNA of cyanophage LPP-3]. / Fizicheskoe kartirovanie DNK tsianofaga LPP-3.

Restrictases fit for the purposes of physical mapping of cyanophage LPP-3 DNA have been selected as a result of the restriction analysis. The use of the methods of mutual hydrolysis, restriction of the fragment isolated from gel and terminal labeling allowed formation a physical map of LPP-3 cyanophage DNA with the complete scheme of allocation of 14 sites for 8 restrictases: Alw44I, Bsp1191, BsuRI, Eco147I, EheI, NcoI, Kpn2I and PvuI as well as the position of certain sites for restrictases HindIII, KpnI and Sau3A.

[Use of cyanobacteria for indication of toxic action of stachybotrys toxins]. / Vikorystannia tsianobakterii dlia indykatsiï toksychnoï diï stakhibotriotoksyniv.

Screening of cyanobacteria cultures for their sensitivity to the action of complex preparations of toxins from 18 strains of Stachybotrys chartarum has been carried out with the purpose to search for new test-organisms to create a reliable system of biotesting of stachibotryotoxines. In contrast to other cultures of unicell (Anacystis nidulans) and filamentous (Anabaena sp., Anabaena variabilis, Nostoc linckia, Plectonema boryanum) cyanobacteria used in the experiment, only one culture of unicell cyanobacterium Synechococcus cedrorum proved to be highly sensitive to the action of preparations of the whole group of studied mycotoxins. Their activity with respect to this culture varied within wide limits (lysis zone diameter is 11-30 mm) and for the most of strains it coincided with or exceeded this index determined with the use of other biotests. Owing to the use of Synechococcus cedrorum as test culture the authors have proved for the first time the toxicity of three strains of Stachybotrys chartarum (K15822, 14722, 14186). Basing on the data obtained the authors have determined the culture of the unicell cyanobacterium, according to the sign of high sensitivity to all the studied stachibotryotoxin preparations of solid and liquid nutrient media, as a new test-organism for detection of this group of toxins and it may be recommended for practical use. Optimal conditions for growing test-culture and biotesting of toxins have been defined.

[Some peculiarities of DNA structure of cyanophage LPP-3]. / Nekotorye osobennosti struktury DNK tsianofaga LPP-3.

The efficiency of radioactive labeling of 3'- and 5'-ends of cyanophage LPP-3 DNA by polynucleotide kinase T4 and terminal transferase under various reaction conditions has been investigated. The obtained data prove that cyanophage LPP-3 DNA has the protruding 3'-ends. The experiments on ligation of native molecules of LPP-3 DNA evidence that the virus genome ends do not display any complimentarity. Separate fragments of LPP-3 DNA were cloned. The restriction analysis of the cloned fragments has confirmed a supposition on the absence of LPP-3 cyanophage of GGGCC and GGCCC sequences in the genome. A hypothesis has been suggested about similar site-specificity of the virus. Counterselection of the genome LPP-3 cyanophage allows it to be considered a promising one in the construction of new cloning vectors in cyanobacterium.

[Development of the method of RNA labeling in vitro using the terminal transferase with the purpose to obtain hybridization probes]. / Razrabotka metoda mecheniia RNK in vitro s pomoshch'iu terminal'noi transferazy s tsel'iu polucheniia gibridizatsionnykh zondov.

A possibility to use terminal transferase to obtain hybridized RNA-probes has been investigated. Optimal conditions of the reaction proceeding were elaborated on the model objects--RNA of the phage MS2: 200 mM of potassium cacodylate, pH 7.2, 1 mM CoCl2, 1 mM DTT, under the considerable surplus of the enzyme. The elaborated method was used to determine the area of early genes of cyanophage LPP-3. With this purpose total RNA preparations were isolated from cyanobacterium Plectonema boryanum CALU 465. The above preparations were labeled by means of terminal transferase and [alpha-32P] ATP and used as the probes in hybridization with the DNA restriction fragments of LPP-3. Proceeding from the results of Southern-blots, a conclusion was made, that the KpnI-fragment of genome of cyanophage LPP3 10 kb in size includes the early genes. A possibility of practical use of the developed method is discussed.

[Comparative characteristics of physico-chemical and regulating properties of aspartate kinases from three species of cyanobacteria]. / Porivnial'na kharakterystyka fizyko-khimichnykh ta rehuliatornykh vlastyvostei aspartatkinazy tr'okh vydiv tsianobakterii.

Aspartokinases have been isolated from the cell-free extracts of Plectonema boryanum, Anabaena variabilis and Synechococcus cedrorum. Their physico-chemical characteristics and peculiarities of retroregulation by amino acids were studied. It has been shown that in P. boryanum cells aspartokinase reaction is catalyzed by only one enzyme, which is similar to the previously described bacterial origin enzymes. Three izoenzymes, which differ in their main properties and amino acids-effectors, have been founded in the cells of A. variabilis. One enzyme with aspartokinase activity, which differs from other cyanobacteria enzymes in some physico-chemical features has been founded in the cells of S. cedrorum.

[Physical mapping of plasmid pSM1 of the cyanobacterium Plectonema boryanum GOM. strain CALU 465]. / Fizicheskoe kartirovanie plazmidy pSM1 iz tsianobakterii Plectonema boryanum GOM. shtamm CALU 465.

A physical map of plasmid pSM1 has been compiled as a result of restriction analysis of pBSMI isolated from the cells of cyanobacterium Plectonema boryanum 465. A comparative analysis of the restriction sites localization in DNA of plasmid pSM1 with site topology of plasmids pGL3 of P. boryanum and pPF1 of Phormidium foveolarum indicates to high degree of these plasmids homology.

[Construction of the shuttle vector pSTS1 and development of the method of its introduction into the cells of the cyanbacterium Plectonema boryanum CALU 465]. / Konstruirovanie chelnochnogo vektora pSTS1 i otrabotka sposoba ego vvedeniia v kletki tsianobakterii Plectonema boryanum GOM. shtamm CALU 465.

It has been shown that cells of trichoma cyanobacterium Plectonema boryanum G o m. strain CALU 465 do not possess natural resistance to antibiotics ampicyllin, chloramphenicol, streptomycin and canamycin. A shuttle vector pSTS1 has been constructed on the basis of cyanobacterial plasmid pSM1 and vector pBR 322. Conditions have been developed and conjugative transfer of pSTS1 to the cells of cyanobacterium P. boryanum has been performed. As the result of expression of beta-lactamase gene pSTS1 the resistance of cells-conjugantes to ampicillin increased by three orders.

[Use of associative cultures of cyanobacteria for tertiary treatment of sewage from yeast and alcohol plants]. / Vykorystannia asotsiatyvnykh kul'tur tsianobakterii dlia ochystky stichnykh vod spyrto-drizhdzhovykh zavodiv.

The associative cultures of cyanobacteria were selected which may be used for development of technology of tertiary treatment of sewage for yeast and alcohol production plants. Growth parameters of associations were studied during periodic and continuous cultivation. When studying the influence of flowing speed for some exponents of treatment it was shown that cultivation of cyanobacteria should be carried out with the change of volume not more than 0.01 h. Final treatment 0.010 h-1 in order to return sewage to natural water reservoirs may be carried out by means of Oscillatoria sp. in bioponds.

[The isolation, purification and cloning of the pSM1 plasmid from the cyanobacterium Plectonema boryanum]. / Vydelenie, ochistka i klonirovanie plazmidy pSM1 iz tsianobakterii Plectonema boryanum.

Three methods of plasmids isolation from the cells of cyanobacteria: Plectonema boryanum 465, P. nostocorum 636, P. nostocorum 638, P. edaphicum 262, Plectonema sp. 456, Phormidium uncinatum 528, Anabaena variabilis 458 and Synechococcus cedrorum 750 have been approved. The method of plasmids isolation by 1 M NaCl salting out was used for plasmid screening. All the strains studied contained large, above 55 KB plasmids, small plasmids (1.5 KB) were found in the cells of P. boryanum 465 and P. nostocorum 636. After the restriction analysis a small plasmid of P. boryanum 465 was chosen to be used in the experiments on the vector construction. The above plasmid was called pSM1 was cloned into the vector pBluescri ptIISK(+) on the unique ClaI site. The obtained construction pBSMI may be used as the basic one for creation of cyanobacterial vectors.

[Activity of tricarboxylic acid cycle enzymes in cyanobacteria Spirulina platensis]. / Izuchenie aktivnosti fermentov tsikla trikarbonovykh kislot tsianobacterii Spirulina platensis.

The activity level and some physico-chemical properties of enzymes of the tricarboxylic acid cycle (TCA cycle) and the associated enzymes isocitrate lyase and glutamate dehydrogenase of cyanobacterium Spirulina platensis grown under illumination of 5000 lk in batch conditions, have been studied. High activities of most of the studied enzymes except for alpha-ketoglutarate dehydrogenase (alpha-KGDH) and succinate dehydrogenase have been estimated. In some cases the activities were by an order higher than that of similar enzymes in other cyanobacteria. This reflects the microorganism ability to synthesize intensively organic substances and first of all protein. Absence of alpha-KGDH activity proves that TCA cycle of spirulina has a limited value for energy generation and mainly performs the biosynthetic function.

[Key enzymes of biosynthesis of amino acids of the glutamic series in the virus-cell system Anabaena variabilis + A-1]. / Kliuchevye fermenty biosinteza aminokislot glutaminovogo riada v virus-kletochnoi sisteme Anabaena variabilis +A-1.

The influence of cyanophage A-1 reproduction on glutamate dehydrogenase (GDG) and glutamine synthetase (GS) in A. variabilis cells was studied. It was determined that the both enzymes are intensified by 70% and 30%, accordingly. Isoenzymes of GDG and GS were isolated from native and infected cells of cyanobacteria, they had various physicochemical properties. It is concluded that cyanophage development causes the specific modification of cell enzymes.

[Phagolysates of cyanobacteria: their biocidal properties and use]. / Fagolizaty tsianobakterii: ikh biotsidnost' i ispol'zovanie.

Data on the biological activity of cyanobacterial phagolysates were obtained in experiments. They concern the organisms of different evolutional level, such as some conventional pathogenic bacteria, plant and root parasitic nematodes and phytophagous insects. The authors suppose the specific mechanism of biocidal and inhibitory action of cyanobacteria and their phagolysates in respect to different living systems. These facts are very important for the elaboration of practical aspects of algal metabolites employment in agriculture and medicine.