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Fig. 2 | Plant Methods

Fig. 2

From: Tagging and catching: rapid isolation and efficient labeling of organelles using the covalent Spy-System in planta

Fig. 2

Transient expression kinetics and covalent intermolecular bond formation between organelle-specific SpyTag and eGFP-SpyCatcher in N. benthamiana. Western Blot analysis of leaf samples transiently co-expressing HA-tagged organelle-specific SpyTag and eGFP-SpyCatcher. Samples were taken every 24 h until 96 h after infiltration (lane 1–5). Lane 1: 0 h, lane 2: 24 h, lane 3: 48 h, lane 4: 72 h and lane 5: 96 h after infiltration. Co-expression and interaction of eGFP-SpyCatcher and a Plastid-SpyTag (plastid-specific; 10 kDa) and b Mito-SpyTag (mitochondrion-specific; 8 kDa) is shown. c The expression of eGFP-SpyCatcher alone (38 kDa, negative control). MW stands for molecular weight (kDa). Black arrows indicate the single protein bands, red arrows show the covalent product after SpyTag/SpyCatcher interaction

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