Fig. 2
Signal peptide validation and subcellular localization of UV_1261. a Protein sequence of UV_1261. Bold and underlined letters represent putative signal peptide. Shaded letters indicate cysteine residues. b Validation of UV_1261 signal peptide by yeast invertase secretion assay. Predicted signal peptide sequence of UV_1261 was fused in-frame to yeast mature invertase sequence in pSUC2 vector and expressed in YTK12. Functional signal peptide could enable yeast growth on both CMD-W and YPRAA. N-terminal sequence of Mg87 and signal peptide of Avr1b were used as the negative and positive control, respectively. c The construct UV_1261-eYFP was transformed into Arabidopsis Col-gl. After obtaining T2 transgenic lines, leaves expressing UV_1261-eYFP were checked under a confocal laser scanning microscope. Nuclei were stained by PI and false-colored in red. Chloroplasts showed autofluorescence and was false-colored in blue. White arrows point to chloroplasts, and white triangles point to nuclei. Scale bar, 20 μm. d Fluorescence intensity curves were drawn according to the direction of the yellow arrow in (c). e Western blot with GFP antibody in Arabidopsis leaves expressing UV_1261-eYFP. The band at 40 kDa indicates the UV_1261-eYFP fusion protein