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Fig. 3 | Plant Methods

Fig. 3

From: A plant protein farnesylation system in prokaryotic cells reveals Arabidopsis AtJ3 produced and farnesylated in E. coli maintains its function of protecting proteins from heat inactivation

Fig. 3

AtJ3 was farnesylated in AtPFT-producing E. coli cells. a AtJ3 produced in E. coli cells harboring either pGEX-AtJ3 alone or both pGEX-AtJ3 and pRSF-Duet1-AtPFT was purified using Glutathione Sepharose 4B beads and run in duplicate, one for Coomassie blue staining (left panel) and one for western blot detection (right panel), using SDS-PAGE. The AtJ3 purified from pGEX-AtJ3 and pRSF-Duet1-AtPFT co-transformed E. coli cells (AtPFT + AtJ3, red arrowhead) exhibited faster electrophoretic mobility than that from cells harboring pGEX-AtJ3 alone (AtJ3, black arrowhead), indicating AtJ3 from co-transformed cells was farnesylated. b The extracted ion chromatograms (EIC) of the C-terminal peptide of AtJ3 with and without farnesylation. The target peptide (AQAQREAYDDDDEDDDHPGGAQRVQC) produced from the cells containing pGEX-AtJ3 alone (upper panel) showed only a non-farnesylated form with 741.0565 (4+) m/z at 18.52 min, but the target peptide produced from the cells containing pRSF-Duet1-AtPFT and pGEX-AtJ3 (lower panel) showed both non-farnesylated form with 741.0565 (4+) m/z at 19.47 min and farnesylated form with 1036.7902 (3+) m/z at 64.52 min, respectively

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Plant Methods

ISSN: 1746-4811