Expression, purification and characterization of recombinant Arabidopsis thaliana cytokinin dehydrogenase AtCKX2
The result's identifiers
Result code in 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>
Result on the web
—
DOI - Digital Object Identifier
—
Alternative languages
Result language
angličtina
Original language name
Expression, purification and characterization of recombinant Arabidopsis thaliana cytokinin dehydrogenase AtCKX2
Original language description
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.
Czech name
Exprese, purifikace a charakterizace rekombinantní cytokinindehydrogenasy AtCKX2 z Arabidopsis thaliana
Czech description
Cytokinin dehydrogenasa byla klonována do expresních plasmidů pYES2 a pDR197 nebo integrována do genomu kvasinky S. cerevisiae pomocí vektorů YIplac211 a pYESX pod konstitutivním nebo indukovatelným promotorem. CKX protein byl ve všech případech sekretován do kultivačního média. AtCKX2 byla purifikována do homogenity z koncentrovaného média klonu pYESX::AtCKX2/ApaI, který produkuje asi 1,5 mg CKX na litr média. Purifikace byla provedena pomocí chromatografie na Concanavalinu A, následované iontově výměnnou, gelovou a hydroxyaptitovou kolonou. AtCKX2 vykazuje pH optimum při pH 7 až 8, reaguje s izoprenoidními i aromatickými cytokininy a jako elektronové akceptory využívá chinony a DCPIP.
Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
CE - Biochemistry
OECD FORD branch
—
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
Z - Vyzkumny zamer (s odkazem do CEZ)
Others
Publication year
2005
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Biologia Plantarum
ISSN
0006-3134
e-ISSN
—
Volume of the periodical
49
Issue of the periodical within the volume
S1
Country of publishing house
CZ - CZECH REPUBLIC
Number of pages
40
Pages from-to
"S4"
UT code for WoS article
—
EID of the result in the Scopus database
—