Naslov
Influence of different gene domains on gene trageting in yeast

Autori
Gjuračić, Krešimir ; Bruschi, C.V.

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

Izvornik
19th International Conference on Yeast Genetics and Molecular Biology ; u: Current Genetics 35 (1999) (S) / - Heidelberg : Springer, 1999, 3-4

Skup
International Conference on Yeast Genetics and Molecular Biology (19 ; 1999)

Mjesto i datum
Rimini, Italija, 25.05.1999. - 30.05.1999

Vrsta sudjelovanja
Pozvano predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
yeast; gene targeting; secondary DNA structure

Sažetak
During meiotic recombination in yeast, intergenic regions are more recombinogenic and within them double-strand breaks (DSBs) are induced. Among these regions there isn't any specific DNA sequence found, therefore we made the working hypothesis that secondary DNA structures might be a signal for the introducing breaks within these regions. In silico analysis of the DNA sequence of the yeast chromosome III (two softwares that predict position of the significant secondary DNA structures were used: Bend-it and Stemloop) showed a significant correlation between position of DSB and predicted DNA curvature. In approximately 70% of cases the break is induced within or in the close proximity of the hypothetically curved and/or looped DNA regions. To test if there is any similarity in distribution of highly recombinogenic sites along the yeast genome, between meiotic and mitotic recombination, a simple experimental system was designed. Integrative plasmid vectors containing three gene domains of URA3 gene (5’ promoter-containing region, partial ORF and 3’ terminator-containing region), the kanMX4 gene as selective marker and the partial ORF of ADE2 gene, were used for transformation of yeast cells. The difference in the frequency of recombination among three functional domains was estimated as the ratio of integration events into a particular gene domain of URA3 gene (white colonies), to integration of plasmid DNA into the ADE2 gene (red colonies). Preliminary data showed that the terminator-containing intergenic region of the URA3 gene, which previously showed significant curvature, is more recombinogenic (up to 10 times) in comparison to other two URA3 gene domains.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA