Functional expression and purification of cytokinin dehydrogenase from Arabidopsis thaliana (AtCKX2) in Saccharomyces cerevisiae

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F07%3A00004716" target="_blank" >RIV/61989592:15310/07:00004716 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Functional expression and purification of cytokinin dehydrogenase from Arabidopsis thaliana (AtCKX2) in Saccharomyces cerevisiae

Original language description

Cytokinin dehydrogenase (CKX) is responsible for regulating the endogenous cytokinin content by oxidative removal of the side chain and seven distinct genes, AtCKX1 to AtCKX7, code for the enzyme in Arabidopsis thaliana. The recombinant enzyme AtCKX2 wasproduced in Saccharomyces cerevisiae after expressing the corresponding gene from a plasmid (pDR197) or following chromosomal integration, under either the constitutive promoter PMA1 or the inducible promoter GAL1. The recombinant protein was purified from yeast culture media using a sequence of chromatographic steps. The purified enzyme had a molecular mass of 61 kDa and a typical flavoprotein spectrum. The specific activity of the enzyme was 87.8 ?kat g-1, with isopentenyladenine as a substrate and 2,3-dimethoxy-5-methyl-pbenzoquinone as an electron acceptor. The pH optimum lay between 7.0 and 8.0, depending on the electron akceptor used. AtCKX2 reacts both with isoprenoid and aromatic cytokinins, the activity with isoprenoid cytokin

Czech name

Funkční exprese a purifikace cytokinin dehydrogenasy z Arabidopsis thaliana (AtCKX2) v Saccharomyces cerevisiae

Czech description

Cytokininoxidasa/dehydrogenasa byla produkována kvasinkou Saccharomyces a rekombinantní protein byl purifikován. Byly posány vlastnosti purifikovaného proteinu: pH optimum, substrátová specifita a preference pro různé elektronové akceptory.

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

EB - Genetics and molecular biology

OECD FORD branch

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Project

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Continuities

Z - Vyzkumny zamer (s odkazem do CEZ)

Publication year

2007

Confidentiality

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

Name of the periodical

Biologia Plantarum

ISSN

0006-3134

e-ISSN

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Volume of the periodical

51

Issue of the periodical within the volume

4

Country of publishing house

CZ - CZECH REPUBLIC

Number of pages

10

Pages from-to

673-682

UT code for WoS article

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EID of the result in the Scopus database

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