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. 2005 Aug 4;102(33):11928–11933. doi: 10.1073/pnas.0505461102

Fig. 1.

Fig. 1.

Generation of epitope-tagged AGO1 transgenic Arabidopsis. (A) Diagram of the FLAG-AGO1 construct. Positions of the restriction sites used for the cloning are given relative to the start codon. The sequence and position of the FLAG epitope is indicated. Thick lines, regions encoding the PAZ and PIWI domains; thin broken arrow, translation start; black dot, translation stop. (B) Diagram of the AGO1 genomic locus. Gray boxes, exons; triangle, T-DNA insertion in ago1 mutant Salk_087076 line (ago1-36) with left border (Lb) and right border (Rb) orientation. Other symbols as in A.(C) FLAG-AGO1 complements the ago1-36 phenotype. Photographs are taken 2 weeks postgermination. (D) PCR genotyping of the FLAG-AGO1 line. The ago1-36 (Upper) and not the WT allele (Lower) is amplified from the selected FLAG-AGO1 transgenic line. (E) Expression of FLAG AGO1 transcripts. ago1-36 mutants produce a truncated transcript comprising the sequence 5′ (Middle) but not 3′ (Bottom) of the T-DNA insertion. Expression of a full-length AGO1 transcript is restored in the selected FLAG-AGO1 line. Actin primers (Top) were used to confirm equal loading, and reactions without reverse transcriptase were performed to exclude DNA contamination. DNA, control PCR with genomic DNA.