Optimized Detection of Protein-Protein and Protein-DNA Interactions, with Particular Application to Plant Telomeres

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F20%3A00604833" target="_blank" >RIV/68081707:_____/20:00604833 - isvavai.cz</a>

Výsledek na webu

<a href="https://link.springer.com/protocol/10.1007/978-1-0716-0763-3_11" target="_blank" >https://link.springer.com/protocol/10.1007/978-1-0716-0763-3_11</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/978-1-0716-0763-3_11" target="_blank" >10.1007/978-1-0716-0763-3_11</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Optimized Detection of Protein-Protein and Protein-DNA Interactions, with Particular Application to Plant Telomeres

Popis výsledku v původním jazyce

Characterization of protein-protein and protein-DNA interactions is critical to understand mechanisms governing the biology of cells. Here we describe optimized methods and their mutual combinations for this purpose: bimolecular fluorescence complementation (BiFC), co-immunoprecipitation (Co-IP), yeast two-hybrid systems (Y2H), and chromatin immunoprecipitation (ChIP). These improved protocols detect trimeric complexes in which two proteins of interest interact indirectly via a protein sandwiched between them. They also allow isolation of low-abundance chromatin proteins and confirmation that proteins of interest are associated with specific DNA sequences, for example telomeric tracts. Here we describe these methods and their application to map interactions of several telomere- and telomerase-associated proteins and to purify a sufficient amount of chromatin from Arabidopsis thaliana for further investigations (e.g., next-generation sequencing, hybridization).

Název v anglickém jazyce

Optimized Detection of Protein-Protein and Protein-DNA Interactions, with Particular Application to Plant Telomeres

Popis výsledku anglicky

Characterization of protein-protein and protein-DNA interactions is critical to understand mechanisms governing the biology of cells. Here we describe optimized methods and their mutual combinations for this purpose: bimolecular fluorescence complementation (BiFC), co-immunoprecipitation (Co-IP), yeast two-hybrid systems (Y2H), and chromatin immunoprecipitation (ChIP). These improved protocols detect trimeric complexes in which two proteins of interest interact indirectly via a protein sandwiched between them. They also allow isolation of low-abundance chromatin proteins and confirmation that proteins of interest are associated with specific DNA sequences, for example telomeric tracts. Here we describe these methods and their application to map interactions of several telomere- and telomerase-associated proteins and to purify a sufficient amount of chromatin from Arabidopsis thaliana for further investigations (e.g., next-generation sequencing, hybridization).

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10609 - Biochemical research methods

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Methods in Molecular Biology

ISSN

1064-3745

e-ISSN

—

Svazek periodika

2175

Číslo periodika v rámci svazku

2021-08-12

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

29

Strana od-do

139-167

Kód UT WoS článku

000681366300012

EID výsledku v databázi Scopus

2-s2.0-85088227850