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Pregled bibliografske jedinice broj: 966407

Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology

Lepur, Adriana; Vugrek, Oliver
Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology // Methods in molecular biology / Clifton, N.J. (ur.).
Totowa (NJ): Springer, 2018. str. 259-267 doi:10.1007/978-1-4939-7871-7_17

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Naslov
Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology

Autori
Lepur, Adriana ; Vugrek, Oliver

Vrsta, podvrsta i kategorija rada
Poglavlja u knjigama, znanstveni

Knjiga
Methods in molecular biology

Urednik/ci
Clifton, N.J.

Izdavač
Springer

Grad
Totowa (NJ)

Godina
2018

Raspon stranica
259-267

ISBN
978-1-4939-7870-0

ISSN
1064-3745

Ključne riječi
BiFC ; Complementation ; Fluorescent microscopy ; Fluorescent protein ; Gateway ; Live cells ; Mammalian ; PPIs ; Protein interactions ; Venus

Sažetak
Bimolecular fluorescence complementation (BiFC) is a powerful and sensitive tool to discover new protein-protein interactions (PPIs). It enables visualization and localization of protein-protein interactions (PPIs) in living cells. The idea behind BiFC is to split a fluorescent protein, for example yellow fluorescent protein (YFP), into two parts that are unable to emit fluorescent signal on their own. Therefore, in order to regain fluorescence the split protein fragments must establish close proximity. This is accomplished by fusing the split fragments to proteins that are postulated to interact, and expressing them in living cells. Subsequently, detection of fluorescence indicates interaction of given proteins. Since complementation is practically irreversible it can capture weak and transient interactions. Using suitable vectors for human protein expression, thus avoiding viral cell transfection, we introduced Gateway-based cloning features to the BiFC system, thereby enabling time efficient vector construction in order to maximize the full potential of the BiFC approach to investigate many protein- protein interactions in a high-throughput fashion. This protocol explains steps in a typical protein-protein interaction survey, from the vector selection, cell transfection, and visualization of the fluorescent signal.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Interdisciplinarne prirodne znanosti, Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)



POVEZANOST RADA

Projekti:
FP7-REGPOT-2012-2013-1

Ustanove:
Institut "Ruđer Bošković", Zagreb

Profili:

Avatar Url Oliver Vugrek (autor)

Avatar Url Adriana Lepur (autor)

Poveznice na cjeloviti tekst rada:

doi link.springer.com

Citiraj ovu publikaciju:

Lepur, Adriana; Vugrek, Oliver
Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology // Methods in molecular biology / Clifton, N.J. (ur.).
Totowa (NJ): Springer, 2018. str. 259-267 doi:10.1007/978-1-4939-7871-7_17
Lepur, A. & Vugrek, O. (2018) Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology. U: Clifton, N. (ur.) Methods in molecular biology. Totowa (NJ), Springer, str. 259-267 doi:10.1007/978-1-4939-7871-7_17.
@inbook{inbook, author = {Lepur, Adriana and Vugrek, Oliver}, editor = {Clifton, N.}, year = {2018}, pages = {259-267}, DOI = {10.1007/978-1-4939-7871-7\_17}, keywords = {BiFC, Complementation, Fluorescent microscopy, Fluorescent protein, Gateway, Live cells, Mammalian, PPIs, Protein interactions, Venus}, doi = {10.1007/978-1-4939-7871-7\_17}, isbn = {978-1-4939-7870-0}, issn = {1064-3745}, title = {Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology}, keyword = {BiFC, Complementation, Fluorescent microscopy, Fluorescent protein, Gateway, Live cells, Mammalian, PPIs, Protein interactions, Venus}, publisher = {Springer}, publisherplace = {Totowa (NJ)} }
@inbook{inbook, author = {Lepur, Adriana and Vugrek, Oliver}, editor = {Clifton, N.}, year = {2018}, pages = {259-267}, DOI = {10.1007/978-1-4939-7871-7\_17}, keywords = {BiFC, Complementation, Fluorescent microscopy, Fluorescent protein, Gateway, Live cells, Mammalian, PPIs, Protein interactions, Venus}, doi = {10.1007/978-1-4939-7871-7\_17}, isbn = {978-1-4939-7870-0}, issn = {1064-3745}, title = {Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology}, keyword = {BiFC, Complementation, Fluorescent microscopy, Fluorescent protein, Gateway, Live cells, Mammalian, PPIs, Protein interactions, Venus}, publisher = {Springer}, publisherplace = {Totowa (NJ)} }

Časopis indeksira:


  • Web of Science Core Collection (WoSCC)
    • Book Citation Index - Science (BKCI-S)
  • Scopus

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