We have made a large scale analysis of prototrophic products of spontaneous and induced mitotic recombination within LYS2 gene of the yeast Saccharomyces. The mutant alleles staying in heterozygote with the wild type allele were uncovered and analysed.Among thirteen lys2 mutations used in the study three had reduced frequencies of mitotic gene conversion. These rarely converting mutations gave a remarkably high proportion of reciprocal events (up to 38%) in pairwise combinations, never seen for any other pair of alleles studied. Two of these mutations are the deletions of large parts of LYS2 gene.The results suggest that mispairing in the region of deletion blocks the hybrid DNA migration and leads to the reduced conversion ability of deletions. Comparison of uncovered alleles ratio in all allele combinations tested lead us to another hypothesis about bidirectional migration of hybrid DNA.

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With the goal of developing non-viral techniques for exogenous gene delivery into mammalian cells, we have studied receptor-mediated gene transfer using complexes of plasmid DNA and galactosylated poly-L-lysine, poly(L-Lys)Gal. To evaluate the optimal parameters for efficient gene transfer into human hepatoma HepG2 cells by the DNA-poly(L-Lys)Gal complexes, the bacterial reporter genes lacZ and cat were used. Examination of the reporter gene expression level showed that the efficiency of DNA delivery into the cells depends on the structure of DNA--poly(L-Lys)Gal complexes formed at various ionic strength values. The efficiency of DNA transfer into the cells also depends on DNA/poly(L-Lys)Gal molar ratio in the complexes. Plasmid vector carrying human apolipoprotein A-I (apoA-I) gene was injected as its complex with poly(L-Lys)Gal into rat tail vein. Some level of ApoA-I was detected in the serum of the injected rats. Also, the human apoA-I-containing plasmid was found to be captured specifically by the rat liver cells and transported into the cell nuclei, where it can persist as an episome-like structure for at least a week. After repeated injections of DNA--poly(L-Lys)Gal complexes, the level of human ApoA-I in rat serum increases, probably, due to accumulation of functional human apoA-I gene in the liver cell nuclei. The data seem to be useful for the development of non-viral approaches to gene therapy of cardiovascular diseases.

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