Exprese, purifikace a charakterizace rekombinantní cytokinindehydrogenasy AtCKX2 z Arabidopsis thaliana

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F05%3A00001816" target="_blank" >RIV/61989592:15310/05:00001816 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Expression, purification and characterization of recombinant Arabidopsis thaliana cytokinin dehydrogenase AtCKX2

Popis výsledku v původním jazyce

Cytokinin dehydrogenase (CKX, EC 1.5.99.12) is a flavoprotein catalyzing irreversible oxidative degradation of plant hormones cytokinins. Plants contain small families of genes coding for different CKX proteins. In Arabidopsis thaliana, seven distinct genes have been identified (AtCKX1-AtCKX7) and six of them have been shown to code for functional proteins. The isolation and purification of individual proteins from plant material is hindered by their simultaneous occurrence and low concentration in plant.AtCKX2 gene was cloned into pYES2 and pDR197 yeast expression plasmids or integrated into yeast genome using vectors YIplac211 and pYESX under constitutive promoter PMA1 and inducible promoter GAL1. The Saccharomyces cerevisiae strain 23344c (MATalphaura3) was used. CKX activity was in all cases released into the growth medium. AtCKX2 was purified to homogeneity from concentrated cell-free medium of the pYESX::AtCKX2/ApaI yeast clone producing about 1.5 mg of CKX per liter of medium.

Název v anglickém jazyce

Expression, purification and characterization of recombinant Arabidopsis thaliana cytokinin dehydrogenase AtCKX2

Popis výsledku anglicky

Cytokinin dehydrogenase (CKX, EC 1.5.99.12) is a flavoprotein catalyzing irreversible oxidative degradation of plant hormones cytokinins. Plants contain small families of genes coding for different CKX proteins. In Arabidopsis thaliana, seven distinct genes have been identified (AtCKX1-AtCKX7) and six of them have been shown to code for functional proteins. The isolation and purification of individual proteins from plant material is hindered by their simultaneous occurrence and low concentration in plant.AtCKX2 gene was cloned into pYES2 and pDR197 yeast expression plasmids or integrated into yeast genome using vectors YIplac211 and pYESX under constitutive promoter PMA1 and inducible promoter GAL1. The Saccharomyces cerevisiae strain 23344c (MATalphaura3) was used. CKX activity was in all cases released into the growth medium. AtCKX2 was purified to homogeneity from concentrated cell-free medium of the pYESX::AtCKX2/ApaI yeast clone producing about 1.5 mg of CKX per liter of medium.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

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

Návaznosti

Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2005

Kód důvěrnosti údajů

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

Název periodika

Biologia Plantarum

ISSN

0006-3134

e-ISSN

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Svazek periodika

49

Číslo periodika v rámci svazku

S1

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

40

Strana od-do

"S4"

Kód UT WoS článku

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EID výsledku v databázi Scopus

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