Naslov
Green fluorescent protein as a tag for the study of cytoskeletal proteins in vivo

Autori
Barišić, Karmela ; Žanić Grubišić, Tihana

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

Izvornik
Kongres hrvatskih biokemičara i molekularnih biologa uz međunarodno sudjelovanje, HB2000, Prohgram i knjiga sažetaka / Floegel, Mirna - Zagreb : Farmaceutsko-biokemijski fakultet Sveučilišta u Zagrebu, 2000

Skup
Kongres hrvatskih biokemičara i molekularnih biologa uz međunarodno sudjelovanje, HB2000

Mjesto i datum
Zagreb, Hrvatska, 13.10.2000. - 15.10.2000

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
Green fluorescent protein (GFP); Dictyostelium discoideum; coronin

Sažetak
Since its first application as an ectopically expressed reporter gene and its subsequent use as fluorescent protein tag, the green fluorescent protein (GFP) from the jellyfish Aequorea victoria has become the tool of choice for visualizing various molecular events in living cells. Experiments based on GFP-tagged proteins are ideally suited for studying cytoskeletal functions. The activity of the cytoskeleton is often described by the reorganization of its spatial structure and the targeted localization of individual cellular components such as proteins and organelles. This reorganization of cytoskeleton can be visualized by the addition of fluorescent tags to cytoskeletal proteins. The activities directly associated with the cytoskeleton such as intracellular transport, cell motility and morphological plasticity can be also brought in focus. GFP-tag has been used for the study of coronin functions in vivo. Coronin is an actin-associated protein. It belongs to the WD-repeat family of proteins. 5'-end of GFP gene was fused at 3'-end of coronin gene and vice versa. The fused genes were cloned into pDBsrH vector that contains blasticidinn S resistance cassette. Dictyostelium discoideum wild type cells and coronin null cells were transformed with recombinant vector by electroporation. The transformants that expressed GFP-tagged coronin were identified by fluorescent microscopy. Direct localization using GFP-tagged coronin has shown that coronin is found in a variety of dynamic F-actin rich structures derived from the cell cortex. In chemotactic cells coronin is recruited to the leading edge.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

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