Naslov
Mammalian genome engineering in the yeast Saccharomyces cerevisiae

Autori
Gjuračić, Krešimir

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

Izvornik
Proceedings/Third Croatian Congress of Microbiology with international participation / Balenović, Mirta - Zagreb : Hrvatsko mikrobiološko društvo, 2004, 76-77

Skup
Third Croatian Congress of Microbiology with international participation

Mjesto i datum
Poreč, Hrvatska, 04.10.2004. - 07.10.2004

Vrsta sudjelovanja
Pozvano predavanje

Vrsta recenzije
Domaća recenzija

Ključne riječi
yeast; mammalian; genome; recombineering

Sažetak
Draft sequences of human, chimp, mouse, rat and dog genomes have been released up today, and the sequencing of several other mammalian genomes is underway. Completion of draft sequences and functional studies of individual components of these sequenced genomes are the next big challenges. Our understanding of gene function and control genetic elements in mammalian genomes partially comes from alignment with known sequences from less complex genomes of other eukaryotes or even prokaryotes. However, the real function of many non-related mammalian DNA sequences came from genetic studies in model organisms. Mouse is probably the best model organism for these types of study, due to established sophisticated transgenic systems based upon murine embryonic stem (ES) cells and gene targeting, which allow the introduction of virtually any genetic modification into the mouse genome. Major limitation of these systems is that they require the production of complex targeting and selection constructs. By using conventional cloning methods with restriction enzymes as major tools, the shape of targeting construct is determined by cleavage site in both cloning vector and genomic DNA, leaving little possibility to design desired genetic modification. This has been overcome by development of the new approaches that make use of the homologous recombination to construct targeting vectors. The yeast Saccharomyces cerevisiae with impressively high frequency of homologous recombination was the first choice microorganism for production of recombinant targeting constructs. Thanks to the inherent yeast recombination machinery, it is possible to clone any desired piece of genomic DNA, using technique known as transformation associated recombination or TAR cloning, and to introduce practically limitless genetic changes within future targeting construct. Recently similar techniques, termed recombineering, were also developed in Escherichia coli using phage-encoded instead of bacterial endogenous recombination proteins for construction of targeting vectors. Taken together, these recombination-related techniques represent the powerful tool for engineering of any sequenced mammalian genome. Their advantages and disadvantages in both microorganisms would be discussed.

Izvorni jezik
Engleski

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