A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F01%3A00004566" target="_blank" >RIV/00216224:14310/01:00004566 - 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

A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production

Popis výsledku v původním jazyce

CKI1, a putative cytokinin receptor of A. thaliana, belongs to a family of sensor hybrid histidine kinases. Here we report a successful production and single-step purification of the CKI1 signal receiver domain. The open reading frame coding for the domain was cloned into an expression vector pET-28a+ and the resulting double His-tag fusion protein was produced in E. coli. Western blot analysis revealed that the fusion protein is present mainly in a soluble fraction of the cell lysate. Single-step purification of the fusion protein was achieved by immobilized metal affinity chromatography. We will report protocol yielding milligram quantities of the recombinant protein in purity higher than 95% from single run of a column. The fusion protein was used to raise polyclonal antibodies in rabbits. Preliminary characterization of the antibodies will be presented.

Název v anglickém jazyce

A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production

Popis výsledku anglicky

CKI1, a putative cytokinin receptor of A. thaliana, belongs to a family of sensor hybrid histidine kinases. Here we report a successful production and single-step purification of the CKI1 signal receiver domain. The open reading frame coding for the domain was cloned into an expression vector pET-28a+ and the resulting double His-tag fusion protein was produced in E. coli. Western blot analysis revealed that the fusion protein is present mainly in a soluble fraction of the cell lysate. Single-step purification of the fusion protein was achieved by immobilized metal affinity chromatography. We will report protocol yielding milligram quantities of the recombinant protein in purity higher than 95% from single run of a column. The fusion protein was used to raise polyclonal antibodies in rabbits. Preliminary characterization of the antibodies will be presented.

Druh

D - Stať ve sborníku

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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Projekt

<a href="/cs/project/LN00A081" target="_blank" >LN00A081: Signální dráhy u rostlin</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2001

Kód důvěrnosti údajů

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

Název statě ve sborníku

17th International Conference on Plant Growth Substances, Brno, Czech Republic, July 1-6, 2001, Abstracts

ISBN

80-7157-518-6

ISSN

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e-ISSN

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Počet stran výsledku

1

Strana od-do

151

Název nakladatele

Mendel University of Agriculture and Forestry, Brno

Místo vydání

Brno

Místo konání akce

Brno

Datum konání akce

1. 1. 2001

Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

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