Naslov
Plasmid integration in yeast : conceptions and misconceptions

Autori
Zgaga, Zoran ; Gjuračić, Krešimir ; Svetec, Ivan-Krešimir ; Mitrikeski, Petar Tomev ; Gregorić, Sandra

Vrsta, podvrsta i kategorija rada
Poglavlja u knjigama, znanstveni

Knjiga
Current Studies of Biotechnology. Volume II : Environment

Urednik/ci
Kniewald, Zlatko

Izdavač
Hrvatsko Društvo za Biotehnologiju

Grad
Zagreb

Godina
2001

Raspon stranica
135-142

ISBN
953-98094-3-6

Ključne riječi
gene targeting, genetic recombination, Saccharomyces cerevisiae

Sažetak
Integration of exogenous DNA into the yeast genome occurs by homologous recombination and is both directed and stimulated by the presence of double-strand breaks (DSBs) in transforming DNA. Repair of the break with or without plasmid integration occurs with equal probability and it is generally assumed that no information is lost or gained during this process. However, some important assumptions of current models are challenged by the results obtained by our group. First, we showed that rare illegitimate integrations may also occur in the presence of homology in the yeast genome and that integration in homology does not always result in reconstruction of the functional allele. Second, the presence of point mutations in double-stranded, but not in single-stranded plasmid, decreased integration in homology. However this antirecombinogenic effect was not proportional to the number of mutations. Third, targeted integrations were strongly suppressed by the presence of short heterologous sequences at the ends of the linearized plasmid. Terminal heterology is not always eliminated during integration and this process can lead to the generation of new alleles. In our experimental system, less then 10% of transformants that repaired DSB present on the replicative plasmid contained the plasmid sequence in their chromosomal DNA. We propose that the plasmid integration can be stimulated by repair processes operating on the hybrid DNA formed during recombination

Izvorni jezik
Engleski

Znanstvena područja
Biologija

Napomena
Rad je odabran medju radovima prezentiranim (pozvanio predavanje) na skupu Scientific Conference with International Participation "Biotechnology and Environment", odrzanom od 19.-22.02.2001., Zagreb, Hrvatska.

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