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Image Search Results
Journal: Journal of Experimental Botany
Article Title: A salt-regulated peptide derived from the CAP superfamily protein negatively regulates salt-stress tolerance in Arabidopsis
doi: 10.1093/jxb/erv263
Figure Lengend Snippet: Identification of AtCAPE1 in Arabidopsis . (A) Deduced amino acid sequence of the PROAtCAPE1 product. The predicted secretion signal peptide is underlined. The CAP domain is shaded in grey. The putative AtCAPE1 peptide is shaded in red. The cleavage site predicted to produce AtCAPE1 is indicated with an arrowhead. The putative cleavage signal motif is double-underlined. (B) LC-MS/MS spectrum of the synthetic AtCAPE1. The y-ion is the C-terminal fragments after peptide bond cleavage while the b-ion is the N-terminal fragments. (C) LC-MS/MS spectrum of the identified AtCAPE1 in Arabidopsis . (D) Schema representing the construct used for constitutive overexpression of enhanced yellow fluorescent protein (eYFP)-tagged PROAtCAPE1. The green box shows the CNYx motif. The putative CAPE is shown in red. The numbers indicate the predicted molecular weight of precursor protein tagged with eYFP (45.7kDa) and the cleaved precursor tagged with eYFP (26.3kDa). (E) Production of the precursor PROAtCAPE1 and the cleaved PROAtCAPE1 in CAPE1ox CNYD and CAPE1ox CNAD transgenic plants, where eYFP was fused to PROAtCAPE1 containing wild type (CNYD) and the mutated (CNAD) junction sequence, respectively. T3 seedlings derived from independent transgenic lines were sampled for western blotting with anti-GFP antibody. Coomassie blue staining was used for protein loading control. The lower panel shows the presence of the T-DNA insertions in the transgenic plants by genomic DNA PCR with the primer pair 35S-F’ and eYFP-R’ shown in (D).
Article Snippet:
Techniques: Sequencing, Liquid Chromatography with Mass Spectroscopy, Construct, Over Expression, Molecular Weight, Transgenic Assay, Derivative Assay, Western Blot, Staining