D 2001

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

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

Basic information

Original name

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

Authors

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

Edition

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

Publisher

Mendel University of Agriculture and Forestry, Brno

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

Genetics and molecular biology

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/01:00004566

Organization unit

Faculty of Science

ISBN

80-7157-518-6
Změněno: 27/3/2002 15:50, prof. RNDr. Břetislav Brzobohatý, CSc.

Abstract

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.

Links

LN00A081, research and development project
Name: Signální dráhy u rostlin