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

D 2001

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

BORKOVCOVÁ, Petra, Jan ZOUHAR, Jan HEJÁTKO a Břetislav BRZOBOHATÝ

Základní údaje

Originální název

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

Autoři

BORKOVCOVÁ, Petra, Jan ZOUHAR, Jan HEJÁTKO a Břetislav BRZOBOHATÝ

Vydání

Brno, 17th International Conference on Plant Growth Substances, Brno, Czech Republic, July 1-6, 2001, Abstracts, s. 151-151, 2001

Nakladatel

Mendel University of Agriculture and Forestry, Brno

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

Genetika a molekulární biologie

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/01:00004566

Organizační jednotka

Přírodovědecká fakulta

ISBN

80-7157-518-6

Změněno: 27. 3. 2002 15:50, prof. RNDr. Břetislav Brzobohatý, CSc.

Anotace

V originále

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.

Návaznosti

LN00A081, projekt VaV

Název: Signální dráhy u rostlin

Citovat

BORKOVCOVÁ, Petra, Jan ZOUHAR, Jan HEJÁTKO a Břetislav BRZOBOHATÝ. A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production. In 17th International Conference on Plant Growth Substances, Brno, Czech Republic, July 1-6, 2001, Abstracts. Brno: Mendel University of Agriculture and Forestry, Brno, 2001, s. 151. ISBN 80-7157-518-6.

@inproceedings{372696,
   author = {Borkovcová, Petra and Zouhar, Jan and Hejátko, Jan and Brzobohatý, Břetislav},
   address = {Brno},
   booktitle = {17th International Conference on Plant Growth Substances, Brno, Czech Republic, July 1-6, 2001, Abstracts},
   language = {eng},
   location = {Brno},
   isbn = {80-7157-518-6},
   pages = {151-151},
   publisher = {Mendel University of Agriculture and Forestry, Brno},
   title = {A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production},
   year = {2001}
}

TY  - JOUR
ID  - 372696
AU  - Borkovcová, Petra - Zouhar, Jan - Hejátko, Jan - Brzobohatý, Břetislav
PY  - 2001
TI  - A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production
PB  - Mendel University of Agriculture and Forestry, Brno
CY  - Brno
SN  - 8071575186
N2  - 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.
ER  -

BORKOVCOVÁ, Petra, Jan ZOUHAR, Jan HEJÁTKO a Břetislav BRZOBOHATÝ. A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production. In \textit{17th International Conference on Plant Growth Substances, Brno, Czech Republic, July 1-6, 2001, Abstracts}. Brno: Mendel University of Agriculture and Forestry, Brno, 2001, s.~151. ISBN~80-7157-518-6.

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