[Association of T3111C polymorphism in 3'-untranslated region of the CLOCK gene with the risk of essential arterial hypertension and coronary artery disease in the Russian population Karelia].

Allele and genotype distributions of the T3111C polymorphism in 3'-untranslated region of the CLOCKgene were examined in the groups of Russian patients with essential arterial hypertension (EAH) and coronary artery disease (CAD), and in control group of Russia residents of the Republic of Karelia. The genotype frequency distributions of the polymorphism examined in the EAH and CAD patients were statistically significantly different from that in the individuals without clinical signs of these diseases. The CC genotype frequency in EAH and CAD males was higher, and in the corresponding females it was lower than in males and females from the control group. Male CC carriers were characterized by a possible increased risk of EAH: OR (95% CI) = 1.42 (0.56; 3.58). Moreover, the presence of the CC genotype in males could increase the risk of CAD: OR (95% CI) = 1.58 (0.63; 3.93).

[Construction and properties of a hybrid operon coding the HBcAG and beta-galactosidase of the hepatitis B virus in Escherichia coli]. / Konstruirovanie i svoistva gibridnogo operona, kodiruiushchego HBcAg virusa gepatita v i veta-galaktozidazy Escherichia coli.

A set of plasmids was constructed, that carries the hybrid operons with an artificial region for translation initiation of the second cistron. The SD-sequence situated close to the termination signal of the previous cistron facilitates the reinitiation of translation. Both HBcAg and beta-galactosidase coding cistrons are functionally active. The analysis of expression efficacy shows: 1) The second cistron possesses its own initiation region; ii) the opportunity of translation reinitiation increases of the protein synthesis level. The correlation between the translation initiation efficacy and the structure of the initiator codon was investigated. AUG and UUG provide comparable protein synthesis levels, AUG being 1.5-3 times more effective. Probably, there exists a different efficacy of recognition of initiator codons by ribosomes for the systems with independent and connected initiation of translation. The influence of mRNA secondary structure in the translation initiation region on expression is discussed.

[Introduction of heterologous epitopes at the N-terminal part of the hepatitis B core protein]. / Vvedenie geterogologichnykh épitopov v N-kontsevuiu chast' core-belka virusa gepatita B.

For investigation of proteins possessing assigned immunological properties, plasmids pPS31-42, pPS1-5, pPS2-17, and pPS1P-30 were constructed encoding the hepatitis B core protein (HBcAg) with N-terminally inserted immunodominant epitopes of preS regions (amino acids 31-36 or 94-105 of preS1, or 133-143 of preS2). Analysis of the hybrid proteins with the use of ELISA and immunoelectron microscopy showed that the insertions did not prevent specific aggregation of the protein molecules, the inserted sequences being exposed on the surface of the particles obtained, and both HBcAg and the corresponding preS determinants were antigenically active.

[Heterologous epitopes in the central part of the hepatitis B virus core protein]. / Geterologichnye épitopy v tsentral'noi chasti core-belka virusa gepatita B.

A set of plasmids was constructed determining synthesis of hybrid proteins with insertions of antigenic determinants of preS1 and preS2 regions of HBV in the middle part of HBcAg. The polypeptides containing the 31-36 or monomeric 94-105 preS1 epitopes were water-soluble and formed particles similar to the 27-nm ones of native HBcAg, possessing antigenic properties of both HBcAg and the inserted epitope, while the hybrids containing 133-143 of preS2 or a trimeric form of the 94-105 preS1 epitope became membrane-bound. Another hybrid encoded by plasmid pPS2M31 contained two regions of HBcAg modification: insertion of amino acids 133-143 a.a. (preS2 region) in the N-terminal part and 31-36 (preS1) in the middle part of the carrier. The immunogenicity of the epitope inserted into the middle part of the HBcAg molecule was an order of magnitude higher than that of the same epitope in the N-part of the protein; the fact might be important for constructing artificial immunogens.

[Effective synthesis of oligo(poly)deoxyribonucleotides using an H-phosphonate method in plastic microcolumn]. / Effektivnyi sintez oligo(poli)dezoksiribonukleotidov H-fosfonatnym metodom v plastikovoi mikrokolonke.

A facile technique of manual oligonucleotide synthesis via H-phosphonate approach is developed. Syntheses carried out in pipette tips with siliconised glasswool filters take 3-3.5 min per cycle with 97-98% yields per condensation. The method was used to synthesize 12-55-mers: T7 and PL promoter regions, gene of the signal peptide of the E. coli OmpA protein, oligonucleotides coding for amino acid sequences 94-105 of preS1- and 133-143 of preS2-regions of hepatitis B virus, hybridisation probes, sequencing primers, oligonucleotides for site-directed mutagenesis, etc.

[Study of the process of thermal aggregation of several representative tobamovirus coat proteins]. / Izuchenie protsessa termicheskoi agregatsii belkov obolochki neskol'kikh predstavitelei tobamovirusov.

The role of the specific region of the tobacco mosaic virus (TMV) coat protein (CP) molecule (called "70A degree-region") in the regulation of ordered and unordered CP aggregation was investigated. CPs of the wild type TMV (strain U1), of temperature sensitive mutant with two amino acid substitutions in the "70A degree-region", and of cucumber virus 3 which is related to TMV but has a completely different structure in the "70A degree-region" were used. With the help of two different tests the processes of temperature-induced unordered aggregation of these three CPs were compared in solutions of different ionic strength and pH. On the basis of the data obtained it was concluded that the "70A-region" represents the most thermolabile region in the TMV CP molecule and that local thermal denaturation of this region results in unordered aggregation, when solution conditions (ionic strength and pH) favor formation of relatively large ordered aggregates (20S-"disks" or helical repolymerized protein).