[Drosophila melanogaster Cell Culture as an Experimental Model to Study Recombination in Wolbachia pipientis].

Wolbachiapipientis is an obligate intracellular endosymbiont that commonly infects arthropods. Comparative genomic studies of Wolbachia reveal traces of numerous events of intergenic and intragenic recombination. The molecular mechanisms of recombination in Wolbachia are not currently known.

[Genotypic Diversity of Wolbachia pipientis in Native and Invasive Harmonia axyridis Pall., 1773 (Coleoptera, Coccinellidae) Populations].

The distribution and variability of reproductive symbiotic Wolbachia pipientis bacteria were studied in seven native and six invasive H. axyridis populations. Wolbachia-infected individuals were found in two invasive and two native populations.

[Identification of potentially invasive species of black flies [Diptera: Simuliidae] from Armenia based on an analysis of variability in the mtDNA barcode of the cox1 gene and chromosomal polymorphism].

Black flies (Diptera, Simuliidae) are well known for their medical, environmental, and veterinary importance. The simuliid fauna of Armenia includes 53 species. A number of dominant species are of ecological importance.

[Transfer of mitochondrial DNA to nuclear genome of cells of passaged cell line of Drosophila virilis].

The variability of the chromosomal fragments of the atp6 mitochondrial gene, which is integrated into chromosomal DNA in the lines of flies of different geographic origins and in the passaged cell lines of D. virilis has been analyzed. We did not reveal any nucleotide variability in this DNA marker among the studied fly lines.

[Mitochondrial genome variation in domesticated sable (Martes zibellina)].

The first comparison of mitochondrial variations in sables from captive and natural populations of the Urals, Central Siberia, Yakutia, Kamchatka, and Japan has been performed. The object of comparative analysis was a 427-bp 5' fragment of the mitochondrial control region, including the D-loop. Two main haplogroups of the sable mitochondrial genome have been found, which provides new data for reconstruction of the spread of the sable over its current range.

[Establishment of a new continuous cell line of Drosophila melanogaster strain infected by the intracellular endosymbiotic bacterium Wolbachia pipientis under natural conditions].

Wolbachia pipientis is an obligately intracellular bacterium infecting a number of arthropod and nematode species. At the body level, Wolbachia infection may cause parthenogenesis, feminization of genetic males, male killing, or cytoplasmic incompatibility; it may also be asymptomatic. Of special interest is DNA transfer from Wolbachia to the host insect genome, which was discovered recently.

[Variability of continuous insect cell lines and their identification].

Continuous insect cell lines make a special object of research in biology. Insect cells in the established lines differ in the number of attributes from both normal differentiated, and embryonic cells. The period of genome destabilization necessarily precedes cell line immortalization.

Poly(N-isopropylacrylamide) copolymer films as vehicles for the sustained delivery of proteins to vascular endothelial cells.

The aim of this study was to establish the capacity of thermoresponsive poly(N-isopropylacrylamide) copolymer films to deliver bioactive concentrations of vascular endothelial growth factor (VEGF165) to human aortic endothelial cells (HAEC) over an extended time period. Films were prepared using a 50:50 (w/w) mixture of non-crosslinkable and crosslinkable copolymers of the following monomer compositions (w/w): 85:15, N-isopropylacrylamide (NiPAAm):N-tert-butylacrylamide (NtBAAm); and 85:13:2 NiPAAm:NtBAAm:acrylamidobenzophenone (ABzPh, crosslinking agent), respectively. After crosslinking by UV irradiation, the ability of films to incorporate a fluorescently labeled carrier protein (FITC-labeled BSA, 1 mg loaded per film), at 4 degrees C, was first established.

Rationalising the design of polymeric thermoresponsive biomaterials.

We investigated the cell adhesion and growth of a series of thermoresponsive copolymers of N-isopropylacrylamide (NIPA) and N-tert-butylacrylamide (NtBA) above their lower critical solubility temperatures (LCST). It was found that cell adhesion and growth on the solvent cast films improved with increasing the NtBA content in the copolymers. The improvement was dependent on cell line.

[Mitochondrial DNA polymorphism in the natural populations of the Drosophila virilis species group].

Restriction fragment length polymorphism (RFLP) analysis has been used to evaluate mitochondrial DNA (mtDNA) variation in 12 sibling species forming the Drosophila virilis species group. The variation thresholds corresponding to the interspecific and interstrain levels have been determined. The results indicate that interspecific hybridization has significantly contributed to the evolutionary history of the virilis species group.

Gypsy group retrotransposon Tv1 from Drosophila virilis.

We have determined the nucleotide sequence of the 6868 bp full-size retrotransposon termed 'Tv1'. Tv1 was isolated from the DNA fraction of extracellular virus-like particles of Drosophila virilis culture cells. Tv1 has the typical structure for a gypsy-group retrotransposon.

[Polymorphism of virus-like particles of retrotransposons in Drosophila and yeast cells].

Data on the molecular arrangement of viruslike particles (VLPs) of yeast and Drosophila retrotransposons are presented. Two methods for identifying VLPs from specific retrotransposon families have been offered. The first method is based on VLPs fractionation by electrophoresis in agarose gel under strictly controlled conditions.

Transient expression assay in a baculovirus system using firefly luciferase gene as a reporter.

Transient gene expression assays were developed to assess the function of the regulatory sequences of baculoviruses Bombyx mori nuclear polyhedrosis virus (BmNPV) and Autographa californica nuclear polyhedrosis virus (AcNPV) in insect cells of Bombyx mori and Spodoptera frugiperda, respectively. DNA sequences encoding luciferase (luc) of the firefly Photinus pyralis was successfully employed in the expression assay as a reporter gene. Recombinant plasmids were constructed containing the luc gene under control of baculovirus-specific or heterologous promoters.

[Phenotypic manifestations of reverse transcriptase activity in yeast cells].

The yeast Saccharomyces cerevisiae and Schizosaccharomyces pombe transformed by plasmids containing retrotransposon from yeast or Drosophila under the control of a strong promoter show the remarkable reverse transcriptase activity. The activity results in the impaired yeast growth and decreased mitotic stability of the plasmids. The phenotypic expression of the reverse transcriptase activity is observed within 30 days.

[Expression of homologous and heterologous retrotransposons in a model system of yeast cells].

Yeast cells Saccharomyces cerevisiae were transformed by the recombinant plasmids containing the Yeast retrotransposon Ty and the Drosophila mobile element gypsy under the control of a strong Yeast promoter. The exogenous Ty-element induces the complete cycle of Ty-retrotransposition including the TyRNA synthesis, formation of virus-like particles, synthesis of all reverse transcriptase intermediates in the virus-like particles with the subsequent circles formation and transposition. The Drosophila mobile element gypsy is capable of inducing the formation of the virus-like particles containing RNA, DNA and proteins of the Ty-retrotransposon only.

Retrotransposon transposition intermediates are encapsidated into virus-like particles.

Virus-like particles (VLPs) possessing reverse transcriptase activity are persistently present in Drosophila melanogaster cultured cells and are formed in yeast induced for transposition. Different retrotransposon transposition intermediates consistent with those expected from the model of reverse transcription pathway of retrotransposon transposition have been detected during the analysis of nucleic acids isolated from VLPs. These data indicate that the act of reverse transcription takes place in VLPs which may be considered as functional intermediates of transposition.

[Expression of Drosophila retrotransposon in yeast].

The recombinant plasmids pPTX and pPGX were constructed, containing the sequences of yeast Ty retrotransposon and Drosophila element mdg4, correspondingly. Transformation of yeast by these plasmids lead to induction of reverse transcriptase activity associated with virus-like particles, containing only the sequences of Ty. The data obtained show that mdg4 is capable of expression in yeast and the products of its expression are used to form the yeast virus-like particles.

[Structure of long terminal repeats of transcriptionally active and inactive copies of the mobile dispersed gene MDG3 in Drosophila melanogaster].

The nucleotide sequences of long terminal repeats (LTRs) and adjacent regions are determined in the transcribed and non-transcribed variants of a mobile dispersed genetic element MDG3. MDG3 is similar to other mdg elements. A 4 bp duplication of the host DNA is generated upon its integration.

[Fine structure of long terminal repeats and stages of reverse transcription of mobile dispersed genes in Drosophila].

In Drosophila melanogaster cultured cells, RNA reverse transcription intermediate forms connected with initiation of minus and plus DNA strand synthesis (minus and plus strong-stop DNA) are detected for mobile dispersed genetic elements MDG1, MDG3 and MDG4 (gypsy). A comparative analysis of intermediate forms has proved that mdg elements pass the same stages of reverse transcription as retroviruses, revealing a complete similarity between intermediate products. It has also been established that these three mdg elements possess a common mechanism of reverse transcription, despite their structural differences.

The steps of reverse transcription of Drosophila mobile dispersed genetic elements and U3-R-U5 structure of their LTRs.

Reverse transcription intermediate forms (minus and plus strong-stop DNA) are detected in Drosophila melanogaster cultured cells for mobile dispersed genetic elements mdg1, mdg3, and mdg4 (gypsy). The mdg elements studied possess a common mechanism of reverse transcription, despite their structural differences, and the comparative analysis of intermediate forms proves that mdg elements pass the same stages of reverse transcription as retroviruses. The length of minus strong-stop DNA that locates the RNA start site coincides with the data obtained from S1 nuclease analysis of transcription initiation.

[Detection of intermediates of RNA reverse transcription for mobile dispersed MDG 1 and MDG 3 genes in Drosophila cells].

Poly(A)+RNA-DNA hybrids complementary to mobile dispersed genetic elements MDG 1 and MDG 3 have been isolated from Drosophila melanogaster culture cells. DNA and RNA in these complexes are represented by perfect hybrids. Three types of single-stranded DNA are detected in these hybrids: full-length DNA molecules of MDG containing one and two long terminal repeats (minus strand) and the long terminal repeat itself (plus strand).