Publication details
A Receiver Domain of CKI1 Sensor Kinase: Expression, Purification and Antibody Production
| Authors | |
|---|---|
| Year of publication | 2001 |
| Type | Article in Proceedings |
| Conference | 17th International Conference on Plant Growth Substances, Brno, Czech Republic, July 1-6, 2001, Abstracts |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| 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. |