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

Result code in 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>
Result on the web
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DOI - Digital Object Identifier
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Result language
angličtina
Original language name
A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production
Original language description
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.
Czech name
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Czech description
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Project
<a href="/en/project/LN00A081" target="_blank" >LN00A081: Signaling pathways in plants</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year
2001
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Article name in the collection
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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Number of pages
1
Pages from-to
151
Publisher name
Mendel University of Agriculture and Forestry, Brno
Place of publication
Brno
Event location
Brno
Event date
Jan 1, 2001
Type of event by nationality
WRD - Celosvětová akce
UT code for WoS article
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