MICHALCOVÁ, Lenka, Radka DOPITOVÁ, Jan HEJÁTKO, Markéta RYVOLOVÁ, Martina VÁLKOVÁ and Jan PREISLER. CE-LIF of Green Fluorescent Fusion Proteins for Investigation of Phosphorylation. In 8 th International Interdisciplinary Meeting on Bioanalysis, CECE 2011. 2011. ISBN 978-80-904959-0-6.
Other formats:   BibTeX LaTeX RIS
Basic information
Original name CE-LIF of Green Fluorescent Fusion Proteins for Investigation of Phosphorylation
Name in Czech Výzkum fosforylace GFP fúzních proteinů pomocí techniky CE-LIF
Authors MICHALCOVÁ, Lenka (203 Czech Republic, belonging to the institution), Radka DOPITOVÁ (203 Czech Republic, belonging to the institution), Jan HEJÁTKO (203 Czech Republic, belonging to the institution), Markéta RYVOLOVÁ (203 Czech Republic, belonging to the institution), Martina VÁLKOVÁ (203 Czech Republic, belonging to the institution) and Jan PREISLER (203 Czech Republic, guarantor, belonging to the institution).
Edition 8 th International Interdisciplinary Meeting on Bioanalysis, CECE 2011, 2011.
Other information
Original language English
Type of outcome Conference abstract
Field of Study 10406 Analytical chemistry
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
RIV identification code RIV/00216224:14740/11:00050124
Organization unit Central European Institute of Technology
ISBN 978-80-904959-0-6
Keywords (in Czech) CE-LIF, fosforylace, AHP proteiny
Keywords in English CE-LIF; phosphorylation; AHP proteins
Tags rivok
Changed by Changed by: Mgr. Nikola Kostlánová, Ph.D., učo 12689. Changed: 2. 4. 2012 14:51.
Abstract
In this study, CE-LIF with 405 nm laser diode was used to study AHP5 fused with GFP. AHP5-GFP fusion proteins overexpressed in E. coli were used for initial optimization of CE-LIF technique because of lower concentrations of the fusion protein produced by plant. The stability of the AHP5-GFP sample was examined; the complex was found to be unstable with GFP being released. The GFP release was not affected by either a long-term storage at -80 and -20 C or the addition of one of the inhibitors (EDTA, PMSF). These results were confirmed by the western blotting. The phosphorylation of the sample AHP5-GFP in vitro is under investigation. Gel filtration was used to obtain mass fractions for better understanding electrophoregrams. The mass fractions were investigated by CE-LIF and Western blotting-LIF and Western blotting.
Abstract (in Czech)
Ke studiu protein AHP5 fúzovaného s GFP byla použita metoda CE-LIF s budícím zdrojem laserovou diodou 405 nm. Fúzní protein AHP5-GFP byl exprimován v E.Coli a byl použit k optimalizaci metody, prototože přirozený výskyt tohoto protein je v rostlinách ve velmi nízkých koncetracích. AHP5-GFP je nestabilní a dochází k uvolňování GFP, tato skutečnost není ovlivněna teplotou skladováním (-20 a -80 C) ani přídavkem inhibitorů (EDTA, PMFS). Výsledky byly potvrzeny pomocí western blottingu. Fosforylace vzorku AHP5-GFP in vitro je nadále zkoumána. Pro lepší interpretaci elektroforegramů byl vzorek podroben gelové filtraci, kde byly získány hmotnostní frakce Ty byly zkoumány metodou CE-LIF a western blottingem
Links
ED1.1.00/02.0068, research and development projectName: CEITEC - central european institute of technology
GA203/09/1025, research and development projectName: Multidetekční platforma pro mikrokolonové separace proteinů a peptidů
Investor: Czech Science Foundation, Standard Projects
MSM0021622415, plan (intention)Name: Molekulární podstata buněčných a tkáňových regulací
Investor: Ministry of Education, Youth and Sports of the CR, Research Intents
PrintDisplayed: 24. 1. 2022 07:33