[Cloning of genes, purification and properties investigation of recombinant DNA ligases from the thermophilic archaeon Pyrococcus abyssi and Methanobacterium thermoautotrophicum].

The genes encoding of DNA ligases from the thermophilic archaeon Pyrococcus abyssi (PabDNA ligase) and Methanobacterium thermoautotrophicum (MthDNA ligase) were cloned and expressed in Escherichia coli. The activity of purified enzymes was studied by ligation of two oligonucleotides, one of which had preformed hairpin structure. In the used system the maximal output of reaction products for both DNA ligases was observed near 70 degrees C that is explained by substrate thermostability.

Fragments of noncoding RNA in plasma of human blood.

Mammalian extracellular fluids, such as plasma of blood or plasma of milk, contain cell-free RNAs. Fragments of ubiquitously expressed mRNAs as well as tumor-specific and viral RNAs have been previously revealed in human plasmas by template-defined RT-PCR. In the present work we aimed to detect major forms of human blood plasma RNAs or to reveal new forms by using two approaches which were independent of context of RNA templates.

Increase of tyrosine hydroxylase and dopamine transporter mRNA levels in ventral tegmental area of male mice under influence of repeated aggression experience.

Tyrosine hydroxylase (TH) and dopamine transporter (DAT) mRNA levels in the ventral tegmental area (VTA) of midbrain were measured by multiplex RT-PCR in male mice with repeated experience of social victories (winners) and social defeats (losers) in 10 daily agonistic confrontations. Two independent experiments revealed enhanced TH and DAT mRNA levels in VTA of the winners in comparison with the losers and controls (animals after 5 days of individual housing). A positive correlation between DAT and TH mRNA levels was shown.

[HLA haplotypes and the predisposition to diffuse toxic goiter].

The results of the family immunogenetic study compared to HLA-typing on the population-genetic level have established a population positive association which was more strong with HLA system regions B and DR (antigens B17, B35, DR5). Haplotype HLA A3, B35, DR5 may be considered as the haplotype of predisposition to diffuse toxic goiter. The strength of the association DT3 with antigens A3, B35, DR5 in the family study was superior to that in the population one.

[The role of mutagenic factors in the development of diffuse toxic goiter].

The role of mutagenic factors in the onset of diffuse toxic goiter (DTG) was studied in 49 families of probands with DTG. The evidence obtained on the order in which the child was born in the proband's family, age of the parents at the time of DTG child delivery, implications of occupational factors in DTG onset does not rule out mutagenic effects in the origin of DTG and thus feasibility of the key gene influence on the disease predisposition.

[Effective expression of genes for interleukin-2 and its mutant analogs in E. coli cells].

Recombinant plasmids were constructed for the efficient expression in E. coli cells of the human interleukin-2 (HIL-2) gene and two its mutant analogues obtained by of chemical-enzymic synthesis and polymerase chain reaction (deletion of 14 C-terminal amino acids and a change of the codon for Trp121 to Phe). The recombinant HIL-2 but not the mutant analogues were shown to be biologically active.

[HLA class I and II antigens and diffuse toxic goiter].

The incidence rate of HLA class I and II antigens has been studied in 168 and 82 patients with diffuse toxic goiter, respectively. Positive association is found for HLA antigens A2, A3, A28, B8, B15, B17, Cw4, DR5; negative one with antigens A24, Bw4, Bw6, Cw1, Cw2, Cw3, Cw5, DR9. Prevalence of antigens B15, B17, Cw4, DR5 is associated with thyrotoxicosis severity, A2 with recurrent toxic goiter.

[Assessment of the parameters of the growth kinetics of an experimental tumor].

A model of the Markov stochastic reproduction and death process is studied as applied to the analysis of time intervals necessary for the experimental tumours to achieve the fixed size. The analysis results have shown nonstationarity of the tumour growth processes during early stages of the tumour development. The presence of such a nonstationarity makes the experimental data interpretation difficult when applying the traditional approach to the tumour cell population kinetics.

[Parametric analysis of the survival of cancer patients based on the results of radiation therapy of cervical cancer patients].

An approach based on the parametric distribution of the life span of tumor-bearing individuals was suggested for the analysis of cancer patients survival. Two groups of patients with cancer of the cervix uteri were identified on the basis of the pre-treatment data (before radiation therapy) on rates of tumor growth. Kinetic curves for tumor growth were constructed using the said distribution procedure as well as the data on survival in each study group.

[Cloning of bacteriophage T5 DNA fragments in the plasmid pBR322. Analysis of recombinant plasmids by the method of bonding with RNA-polymerase from Escherichia coli on nitrocellulose filters].

DNA of bacteriophage T5 was hydrolyzed with restriction endonucleases HindIII and BamHI, and subjected to the combined hydrolysis with BamHI+EcoRI and BamHI+ +HindIII. Fragments obtained were cloned in the plasmid pBR322. About 17% of T5 genome were recovered in recombinant plasmids.

Cloning of genes for bacteriophage T5 stable RNAs.

One EcoRI-generated fragment (440 basepairs) and two EcoRI/HindIII fragments (220 and 960 basepairs) from the deletion region of T5 phage have been inserted into the phage lambda XIII and the plasmid pBR322 as vectors. Recombinant DNA molecules were studied by hybridization with in vivo 32P-labeled T5 4-5 S RNAs on nitrocellulose filters. Two-dimensional polyacrylamide gel electrophoretic fractionation and fingerprint analysis of the RNAs eluted from the filters were carried out to identify RNAs coded by cloned fragments.

Cloning of bacteriophage T5 promoters.

Bacteriophage T5 was subjected to combined hydrolysis with the restriction endonucleases PstI and HindIII and the resulting fragments were inserted into the plasmid pBR322. Selection of transformants for Aps-Tcr-phenotype made it possible to screen the hybrid plasmids that contained promoter sequences in the cloned fragments. Two PstI/HindIII fragments, 720 bp (51% of the T5 DNA length) and 1,200 bp (70%) were cloned in this study.