Revista de bioingeniería y ciencia biomédica

Development of Methods for Safe Application of Viral Vectors for GeneEngineering Manipulations

Abstract

Sainova IV*, Valkova IP, Markova TZ and Nikolova EB 

Additional oncogene copy was transferred in normal cells, and additional tumor-suppression gene copy-in malignant cells, respectively, by transfection with the appropriate recombinant DNA-constructs, containing each one the respective gene. The results obtained were compared with the data from insertion of copy from the respective gene in prokaryotic bacteria strains E. coli, by analogical transfection with appropriate gene constructs. In application of low initial infections (high initial dilutions of viral suspensions, respectively) of eventually attenuated by many passages vaccine strains from the heterologous for mammals and mammalian cells avian viral species, only slight ultra-structural changes and signs of partially-formed immature viral particles in the cells, were established, but no cytopathic effect (CPE was observed). In freezing of the so inoculated mammalian cells after addition of cryoprotector Dimethylsulfoxide (DMSO), thawing and re-incubation in fresh cultivation medium, signs of activated cell proliferation were noted. One of the possible explanations could be eventual transfer of nucleotide sequences from viral particles to separate cells because of activated fusion processes on the influence of DMSO in drastic temperature changes. The results obtained proposed a possibility about application of the described methods as available alternatives for both vaccine production and gene-engineering manipulations with cells, compared with the procedures of preparation and application of designed quite expensive recombinant DNA-constructs. Future studies are necessary in this direction.