Naslov
Linearized integrative single-stranded plasmids in yeast transformation

Autori
Gjuračić, Krešimir ; Alačević, Marija ; Zgaga, Zoran

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

Izvornik
Annual Meeting of Croatian Biochemists HB93, Book of abstracts / - Zagreb, 1993

Skup
Annual Meeting of Croatian Biochemists HB93

Mjesto i datum
Zagreb, Hrvatska, 17.06.1993. - 18.06.1993

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
yeast; transformation; single-stranded DNA

Sažetak
We used the single-stranded nonreplicative plasmid pOM that was linearized with restriction endonuclease EcoRV to transform yeast protoplasts. Linearization was achieved after hybridization of the plasmidic DNA with appropriate 17-mer and was confirmed by both agarose gel electrophoresis and transformation of the bacterium Escherichia coli. Circular and linearized plasmids were shown to transform yeast with comparable efficiencies, but the spectrum of transformants obtained in these two cases was remarkably different. Namely, only 3/30 transformants analyzed by Southern blot hybridization were due to the plasmid integration by homologous recombination, while more than 30% of transformants were produced by conversion of the Ty element (6, 1 kb) that was present in the ura3 gene. Additional experiments revealed that the large majority of transformation events resulted from non-specific (or less-specific) plasmid integrations. A possible role of single-stranded tails in homologous recombination will be discussed.

Izvorni jezik
Engleski

POVEZANOST RADA