Intron-tagged epitope: a tool for facile detection and purification of proteins expressed in Agrobacterium-transformed plant cells.
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 beta-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.
Ferrando
A
A
Max-Planck Institut für Züchtungsforschung, Köln, Germany.
Farràs
R
R
Jásik
J
J
Schell
J
J
Koncz
C
C
D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins; D015153:Blotting, Western; D004269:DNA, Bacterial / Q000235:genetics; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D017930:Genes, Reporter; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D007438:Introns; D010940:Plant Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012231:Rhizobium / Q000235:genetics*; D014170:Transformation, Genetic
C120363:ASK1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D004269:DNA, Bacterial; D000939:Epitopes; D010940:Plant Proteins; C036483:T-DNA; D005966:Glucuronidase