[Expression of a synthetic gene for human interleukin-4 in E. coli cells. Preparation of a biologically active protein]. / Ekspressiia sinteticheskogo gena interleikina-4 cheloveka v kletkakh E. coli. Poluchenie biologicheski aktivnogo belka.

Expression E. coli plasmid were constructed in which the human interleukin-4 (hIL4) synthetic gene is controlled by tac promoter. The expression level of the gene depends on the distance between RBS and the initial codon ATG, with the maximal production in case of the nine base pair distance. The recombinant protein, accumulated in the inclusion bodies, was solubilized, renaturated, and purified to homogeneous, biologically active preparation, the yield being 2 mg/g wet cells.

[Isolation of recombinant interleukin-3 produced by E. coli]. / Vydelenie rekombinantnogo interleikina-3, produtsiruemogo E. coli.

A synthetic gene coding for human interleukin-3 (hIL3) was cloned in the plasmid pTE2IL3, the gene expression being controlled by the phage fd PVIII promotor and the phage T7 gene 10 translational enhancer. Under constitutive biosynthesis conditions in E. coli, the accumulation of recombinant hIL3 (in the inclusion bodies) was up to 30-40% of the total cell protein. An effective procedure of the hIL3 isolation is suggested. The hIL3 was solubilized in 5 M guanidinium chloride, renaturated and purified to homogeneity by a single chromatographic step. The protein's yield was 34 mg/g wet cells. The isolated hIL3 showed a specific biological activity.

Dialect-I, species-specific repeated DNA sequence from barley, Hordeum vulgare.

Dialect-1, species-specific repetitive DNA sequence of barley Hordeum vulgare, was cloned and analysed by Southern blot and in situ hybridization. Dialect-1 is dispersed through all barley chromosomes with copy number 5,000 per genome. Two DNA fragments related to Dialect-1 were revealed in λ phage library, subcloned and mapped. All three clones are structurally heterogenous and it is suggested that the full-length genomic repeat encompassing Dialect-1 is large in size. The Dialect-1 DNA repeat is represented in the genomes of H. vulgare and ssp. agriocrithon and spontaneum in similar form and copy number; it is present in rearranged form with reduced copy number in the genomes of H. bulbosum and H. murinum, and it is absent from genomes of several wild barley species as well as from genomes of wheat, rye, oats and maize. Dialect-1 repeat may be used as a molecular marker in taxonomic studies and for identification of barley chromosomes in interspecies hybrids.

[Direct and reverse transport of RNA in a system of isolated HeLa cell nuclei]. / Priamoi i obratnyi transport RNK v sisteme s izolirovannymi iadrami kletok HeLa.

In a system containing isolated HeLa cell nuclei the release of RNA from the nuclei may be paralleled with the antagonistic process, i. e., RNA translocation into the nuclei. The RNA release from the nuclei depends on incubation time, pH, Mg2+ and nucleoside triphosphate concentration. The rate of reverse transport depends on pH, size of RNA to be translocated and the physiological state of the nuclear membrane. Low molecular weight RNAs (less than 10 S) are translocated into the nuclei most effectively. The nuclei of synchronized HeLa cells in the G1-period are more "permeable" for translocated RNA as compared with the S-phase HeLa cell nuclei.

[RNA transport from isolated nuclei of cultured Djungarian hamster cells]. / Izuchenie transporta RNK is izolirovannykh iader kul'tiviruemykh kletok dzhungarskogo khomiachka.

The in vitro system of RNA transport containing isolated nuclei of Djungarian hamster cells transformed by SV-40 virus was studied. A functional test for cytoplasmic contaminations of the nuclei was proposed. The release of the newly synthesized RNA was found to be dependent on the duration of incubation, temperature and pH of the incubation medium as well as on the presence of spermine, spermidine, dithiothreitol, Mg2+, EDTA, exogenous RNA, nucleoside triphosphates and cytosol. The results obtained indicate that RNA release is an active process with activation energy of 12 kcal/mol. ATP, GTP, CTP and UTP have equal ability to serve as energy sources for the release of RNA. The nucleoside triphosphatase activity of the nuclei was the same in the presence of these four nucleoside triphosphates.

[Kinetic regularities of RNA synthesis. VI. Reverse problem of RNA accumulation in synchronous cell cultures]. / Kineticheskie zakonomernosti sinteza RNK. VI. Obratnaia zadacha kinetiki nakopleniia RNK v sinkhronnoi kletochnoi kul'ture

An algorithm is presented for determining relative nucleotide composition and rates of synthesis of some RNA fractions according to the experimental data of the change with time of mean nucleotide composition of the mixture of RNA fractions of synchroneous cell culture. Piece-linear models with voluntary number of fractures are taken as the models of accumulation of separate RNA types.