02666nam a2200301Ia 4500 MX-MdCICY 20260521091143.0 CICY B-9508 Gateway-compatible vectors for plant functional genomics and proteomics The Plant Journal, 45, p.616-629, 2006 Gateway cloning technology facilitates high-throughput cloning of target sequences by making use of the bacteriophage lambda site-specific recombination system. Target sequences are first captured in a commercially available 'entry vector' and are then recombined into various 'destination vectors' for expression indifferent experimental organisms. Gateway technology has been embraced by a number of plant aboratories that have engineered destination vectors for promoter specificity analyses, protein localization studies, protein/protein interaction studies, constitutive or inducible protein expression studies, gene knockdown by RNA interference, or affinity purification experiments. We review the various types of Gateway destination vectors that are currently available to the plant research community and provide links and references to enable additional information to be obtained concerning these vectors. We also describe a set of 'pEarleyGate' plasmid vectors for Agrobacterium-mediated plant transformation that translationally fuse FLAG, HA, cMyc, AcV5 or tandem affinity purification epitope tags onto target proteins, with or without an adjacent fluorescent protein. The oligopeptide epitope tags allow the affinity purification, immunolocalization or immunoprecipitation of recombinant proteins expressed in vivo. We demonstrate the utility of pEarleyGate destination vectors for the expression of epitope-tagged proteins that can be affinity captured or localized by immunofluorescence microscopy. Antibodies detecting the FLAG, HA, cMyc and AcV5 tags show relatively little cross-reaction with endogenous proteins in a variety of monocotyledonous and dicotyledonous plants, suggesting broad utility for the tags and vectors. AFFINITY PURIFICATION EPITOPE TAG FUSION PROTEIN PROTEIN LOCALIZATION RECOMBINATIONAL CLONING Earley, K.W. Haag, J.R. Pontes, O. Opper, K. Juehne, T. Song, K. Pikaar, C.S. https://drive.google.com/file/d/1mbD8cpQrxEDt9o4hqFLZ94wgP-47Ftzl/view?usp=drivesdk Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx Loc REF1 250602s9999 xx |||||s2 |||| ||und|d 19697 19697 0 0 Loc 0 0 F1 CICY CICY RE 2025-06-25 0 B-9508 2025-06-25 14:06:14 2025-06-25 REF1