Intron-tagged epitope: a tool for facile detection and purification of proteins expressed in Agrobacterium-transformed plant cells
Article first published online: 25 DEC 2001
The Plant Journal
Volume 22, Issue 6, pages 553–560, June 2000
How to Cite
Ferrando, A., Farràs, R., Jásik, J., Schell, J. and Koncz, C. (2000), Intron-tagged epitope: a tool for facile detection and purification of proteins expressed in Agrobacterium-transformed plant cells. The Plant Journal, 22: 553–560. doi: 10.1046/j.1365-313x.2000.00763.x
- Issue published online: 25 DEC 2001
- Article first published online: 25 DEC 2001
- Received 10 January 2000; revised 13 March 2000; accepted 20 March 2000.
Epitope tagging provides a useful tool for immunological detection and cellular localization of proteins in vivo. Using T-DNA-mediated transformation, the detection of epitope-tagged proteins in planta is currently feasible only in transgenic plants, because an artificial expression of cDNA and gene constructs driven by plant promoters in bacteria obscures an early detection of epitope-tagged proteins in Agrobacterium-infected plant cells. We have developed a method for labelling plant coding sequences with intron-tagged epitope-coding domains that are not processed in Agrobacterium. Here we show that the expression of HA-epitope-tagged constructs encoding β-glucuronidase and S-phase kinase-associated (AtSKP1/ASK1) proteins can be specifically and exclusively detected in cultured Arabidopsis cells as early as five days after Agrobacterium infection. This epitope-tagging approach offers an unlimited source of transformed material for purification and localization of proteins expressed individually or simultaneously in Agrobacterium-transformed plant cells.