Naslov
The efficiency of gene replacement in yeast is allele-specific

Autori
Štafa, Anamarija ; Miklenić, Marina ; Lisnić, Berislav ; Svetec, Ivan-Krešimir Svetec ; Zgaga, Zoran

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
EMBO Young Scientist Forum / - Zagreb, 2009

Skup
EMBO Young Scientist Forum

Mjesto i datum
Zagreb, Hrvatska, 15.06.2009. - 17.06.2009

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
gene targeting; gene duplication; aneuploidy; homologous recombination
(gene targeting; gene duplication; aneuploidy; homologous recombination;)

Sažetak
Introduction of linear, non replicative DNA fragments is widely used for the replacement of homologous sequences in the yeast genome. This procedure is very useful for the construction of different yeast strains but is also studied as a general model for targeted genetic manipulations by the "ends-out" recombination. It is generally assumed that, in contrast to higher eukaryotes, transformation of the yeast Saccharomyces cerevisiae with non-replicative DNA occurs almost exclusively by homologous recombination. However, we found out that the ends-out recombination may result in different genetic alterations in the yeast genome and we performed a systematic study of genetic events that may arise as side-effects of gene replacement. In our experimental systems, different URA3 alleles were present on the transforming DNA and in the yeast genome while the spectra of transformation events were determined by Southern blot hybridization. Depending on the experimental system targeted gene replacement ranged from only 60 % to 98 %. Molecular analysis of transformants revealed versatile aberrant recombination events that included illegitimate integration of the transforming DNA, gene or whole chromosome duplications and chromosomal rearrangements. We show that the fidelity of gene targeting in yeast is mainly influenced by the nature of alleles involved in recombination. Additionally, our results suggest that the ends-out recombination involving endogenous DNA fragments may have an important role in the evolution of the yeast genome.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija

POVEZANOST RADA