engleski

Yeast transformation with non-replicative plasmids: different genetic alterations and the choice of the vector

The efficiency of yeast transformation with non-replicative plasmids, as well as the type of genetic alteration introduced, can be significantly influenced by different in vitro modifications of the plasmid. The effects of conformation of the plasmid molecule (double- or single-stranded form), plasmid linearization with restriction enzymes, DNA divergence between chromosomal and plasmidic sequences and UV-irradiation of the plasmid have been studied. These investigations can help us to better understand the mechanisms of genetic recombination but they might also be useful for improving the techniques of yeast strain construction. Introduction of foreign DNA fragments into the yeast genome by illegitimate integration of single-stranded plasmids or increasing of transformation efficiency by UV-irradiation of the plasmid are some of the possible applications of these investigations.

yeast; transformation; homologous recombination; illegitimate recombination; UV irradiation

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