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Pregled bibliografske jedinice broj: 337467

The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells


Deželjin, Martina; Herak Bosnar, Maja; Dubravčić, Klara; Bago, Ružica; Pavelić, Jasminka
The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells // Periodicum biologorum, 110 (2008), 1; 73-76 (međunarodna recenzija, članak, znanstveni)


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Naslov
The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells

Autori
Deželjin, Martina ; Herak Bosnar, Maja ; Dubravčić, Klara ; Bago, Ružica ; Pavelić, Jasminka

Izvornik
Periodicum biologorum (0031-5362) 110 (2008), 1; 73-76

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
GFP ; flow cytometry ; cell-cycle analysis ; adherent cells ; transfection
(The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells)

Sažetak
Background and purpose: GFP (green fluorescent protein) is widely used in a variety of fluorescent methods aimed at revealing the fate of proteins in the cell, intracellular transport, transfection efficiency and is also recommended for cell cycle analysis purposes. In our attempt to evaluate the role of nm23 genes in proliferation of head and neck tumor cells in culture we have decided to use EGFP reporter system and analyze the DNA content by flow cytometry. Materials and methods: To optimize the method we either transiently transfected the cells with pEGFPC1-nm23 constructs or cotransfected the cells with an nm23 carrying constructs and pEGFPC1 as a reporter system. We also established stable clones with pEGFPC1-nm23 constructs and analyzed them by flow cytometry, as well. Results and conclusions: We report our experience for the use of pEGFP reporter system and flow cytometry for determining cell-cycle distribution of transiently and stably transfected adherent tumor cells. We discuss, in brief, the protocol we used and the problems that appeared during our experiments – GFP bleaching, cell clumping and degradation and insufficient number of cells to be analyzed. In conclusion, we suggest useful tips how to avoid or minimize the technical problems of this method and improve the results and analysis.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti



POVEZANOST RADA


Projekti:
065-0982464-2532 - Molekularni mehanizmi nastanka prekanceroznih i kanceroznih lezija usne šupljine (Mravak-Stipetić, Marinka, MZOS ) ( POIROT)
098-0982464-2394 - Gensko liječenje tumora djelovanjem na molekule imunološkog sustava (Pavelić, Jasminka, MZOS ) ( POIROT)
098-0982464-2513 - Uloga gena nm23 u karcinomu pločastog epitela usne šupljine (Herak-Bosnar, Maja, MZOS ) ( POIROT)

Ustanove:
Institut "Ruđer Bošković", Zagreb,
Klinički bolnički centar Zagreb

Poveznice na cjeloviti tekst rada:

Hrčak

Citiraj ovu publikaciju:

Deželjin, Martina; Herak Bosnar, Maja; Dubravčić, Klara; Bago, Ružica; Pavelić, Jasminka
The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells // Periodicum biologorum, 110 (2008), 1; 73-76 (međunarodna recenzija, članak, znanstveni)
Deželjin, M., Herak Bosnar, M., Dubravčić, K., Bago, R. & Pavelić, J. (2008) The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells. Periodicum biologorum, 110 (1), 73-76.
@article{article, author = {De\v{z}eljin, Martina and Herak Bosnar, Maja and Dubrav\v{c}i\'{c}, Klara and Bago, Ru\v{z}ica and Paveli\'{c}, Jasminka}, year = {2008}, pages = {73-76}, keywords = {GFP, flow cytometry, cell-cycle analysis, adherent cells, transfection}, journal = {Periodicum biologorum}, volume = {110}, number = {1}, issn = {0031-5362}, title = {The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells}, keyword = {GFP, flow cytometry, cell-cycle analysis, adherent cells, transfection} }
@article{article, author = {De\v{z}eljin, Martina and Herak Bosnar, Maja and Dubrav\v{c}i\'{c}, Klara and Bago, Ru\v{z}ica and Paveli\'{c}, Jasminka}, year = {2008}, pages = {73-76}, keywords = {The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells}, journal = {Periodicum biologorum}, volume = {110}, number = {1}, issn = {0031-5362}, title = {The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells}, keyword = {The utilization of pEGFP reporter system in cell-cycle analysis of adherent cells} }

Časopis indeksira:


  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus





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