4. Laboratory of Genome Structure.

(1) Staff:

(2) Subject of Research:

(3) General Results 1993-1995:

New original methods of the synthesis of nucleoside 5'-triphosphate derivatives bearing the residues of various fluorescent dyes and photoreagents have been fulfilled and the compounds obtained have been studied as substrates for different DNA and RNA polymerases.
A model for the DNA polymerase translocation along the DNA has been proposed. According to the model, the movement of the enzyme is a result of transition of the enzyme-bound DNA from A- to B-form which is accompanied by the lenghtening of DNA within the binding channel.
An original method of preparation of nonradioactively labelled DNA and RNA probes for molecular hybridization has been suggested. The method is based on incorporation of amino groups into polynucleotides by alkylation and transamination of nucleic base followed by attachment of different labels to these amino groups. The level of labelling of such probes by biotin and fluorochroms which gives maximal sensitivity during dot-, blot- and in situ hybridization has been determined.
The use of high sensitive biotinilated DNA probes allows to map the human horionic somatomammotropin gene to the 17q22 region.
The new method of the DNA photoimmobilization onto the different solid supports which allows to increase the hybridization sensitivity have been developed.
The test-systems for nonradioactive DNA diagnostics of bovine leukemia provirus, Malaria in human blood, and mycoplasmic contamination of cell cultures have been developed.
It has been shown that nuclear proteoglicans are very conservative in mammalian cells. The oligo RNAs from the nuclear proteoglican fraction are able to suppress a transcriptional process in cell culture.
The ISIAH (inherited stress-induced arterial hypertension) rat strain has been characterized by DNA fingerprinting method, and very high genetic homogeneity has been shown.
It has been carried out the mapping of point mutations in leader sequence of the hsp70 gene of various rat strains with hypertensive and normotensive status. The ISIAH rat strain has been shown to carry the point mutation in the leader sequence at a distance of 35 bp from the start of the transcription origin in the recognition site of restrictase BamHI.
The molecular-genetic differences between the fertile and sterile types of cytoplasm in sugar beet have been revealed. The mechanism of spontaneous conversion of normal cytoplasm to sterile one has been suggested.
It has been demonstrated that the rearrangements in mitochondrial DNA appeared during in vitro cultivation of plant cells, part of them has been inherited by regenerated plants. The data have been obtained providing the hypothesis that the higher generation of free oxygen radicals under stress conditions of culturing may be a cause of somaclonal variation.

(4) Publications:

  1. Dikalova A.E., Dudareva N.A., Kubalakova M., Salganik R.I. (1993). Rearrangements in the sugar beet mitochondrial DNA induced by cell suspension, callus cultures and regeneration. Theor.Appl.Genet. 86:699-704.
  2. Popov A.N., Adarichev V.A., Kalachikov S.M., Mishina E.S., Dymshits G.M. (1993). Detection of bovine leukemia provirus by polimerase chain reaction followed by nonradioactive blot hybridization. Problems of virology 38:113-116. (Rus.)
  3. Vlasova I.E., Vlasov V.V. (1993). Molecular evolution: design of nucleic acids possessing catalytic properties and capable of specific complex formation. Molecular Biology 27:1-5.
  4. Adarichev V.A., Dymshits G.M., Mishina E.S., Salganik R.I., Sokhanenkova T.L. Nonradioactive hybridization test onto the Plasmodium falciparum invasion. Biopolymers and cells 9:19-23. (Rus.)
  5. Wlassoff W.A., Kalachikov S.M, Dymshits G.M. (1994). Synthesis of fluorescent labelled uridine 5'-triphosphate derivatives for preparation of nonradioactive RNA probes. Bioorg. Chem. 20:676-681. (Rus.)
  6. Kalachikov S.M., Kel' A.E., Dymshits G.M., Ishov A.M., Borkhsenius S.N. (1994). Cloning of universal hybridization probes for testing mycoplasmal contamination in cell cultures. Molecular Biology 28:298-302.
  7. Vlassova I.E., Nechaeva M.V., Vlassov V.V. (1994). Systems of delivering nucleic acids to cells of mammals. Achievments of modern biology 114:715-726. (Rus.)
  8. Zamaraev K.I., Salganik R.I., Romannikov V.N., Wlassoff W.A., Khramtsov V.V. (1995). The modelling of prebiotic synthesis of oligopeptides in the presence of zeolites and kaolin. Rep. of Russ. Acad. Sci. 340:779- 782. (Rus.)
  9. Wlassoff W.A., Dobrikov M.I., Safronov I.V., Dudko R.Y., Bogachev V.S., Kandaurova V.V., Shishkin G.V., Dymshits G.M., Lavrik O.I. (1995). Synthesis and characterization of (d)NTP derivatives substituted with residues of different photoreagents. Bioconjugate Chemistry 6:352- 360.
  10. Adarichev V.A., Vorob'eva N.V., Grafodatsky A.S., Dymshits G.M., Sablina O.V. (1995). DNA modification with 4-aminohydroxybutylamine to obtain highly sensitive hybridization probes. Location of human chorionic somatomammotropin gene. Molecular Biology 29:307-311.
  11. . Dikalova A.E., Veprev S.G., Maletsky S.I., Salganik R.I. (1995). Polymorphism of mitochondrial genome structure in plants of Beta vulgaris L. with normal cytoplasms. Rep. of Russ. Acad. Sci. (Rus., in press)

(5) Participation in State and International Programs:

(6) Subjects of Scientific Collaboration: