Figure 2
Generation of pEpi::mTRQ2-merC-SYP121 transgenic Arabidopsis plants. (a) Schematic structure of the T-DNA region in the binary vector constructed for generating pEpi transgenic plants. RB, right border; LB, left border; pEpi, putative promoter region of At5g43030; mTRQ2, monomeric variant of cyan fluorescent protein (CFP); merC, bacterial mercury transporter gene; SYP121, plant SNARE gene; t-nos, NOS terminator; Hygr, hygromycin resistance gene. (b) Detection of the T-DNA fragment by PCR from genomic DNA of Col-0 (wild-type) and the pEpi transgenic plants. AtEF1a served as a reference gene. The full-length gel is presented in Supplementary Fig. S1. (c) Quantification of the transgene expression by real-time RT-PCR. Total RNA was extracted from roots and shoots of Col-0 (wild-type), p35S (merC-SYP121 over-expressing line) and the pEpi lines for cDNA synthesis and subsequent real-time RT-PCR analysis. Expression levels of merC were normalized to those of AtEF1a. The data are presented as means with standard deviation (n = 4). Means sharing the same letter are not significantly different (P < 0.05, Tukey’s HSD). (d) Phenotypes of Col-0 (wild-type), p35S (merC-SYP121 over-expressing line) and the pEpi lines grown on the agar plates. A scale bar = 1 cm.
