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Yeast Transformation with Non-Replicative Plasmids: Different Genetic Alterations and the Choice of the Vector

Krešimir Gjuračić*, Milena Ninković**, P. Koren, Marija Alačević and Zoran Zgaga


Faculty of Food Technology and Biotechnology, Laboratory of Biology and Microbial Genetics, Kršnjavoga 25, 41000 Zagreb, Croatia

*Pliva Research Institute, Laboratory of Molecular Biology, Prilaz baruna Filipovića 89, 41000 Zagreb, Croatia

**Institute for Genetic Engineering and Biotechnology, Filipa Kljajića 12, 71000 Sarajevo, Bosnia and Herzegovina

Article history:

Received October 10, 1994

Accepted December 22, 1994

Summary:

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. Introducing 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.

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