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. 2013 Apr 3;8(4):e60801. doi: 10.1371/journal.pone.0060801

Figure 1. Construction of transgenic plants overexpressing A. niger phyA or E. coli appA.

Figure 1

A and B: Schematic representation of binary vector pBI121-phyA (A) and pBI121-appA (B) for B. napus plant transformation. Phytase genes (A. niger phyA and E. coli appA) under the control of the CaMV 35S promoter and nos-terminator were modified for extracellular secretion by inclusion of an extracellular targeting sequence from the carrot extensin (ex) gene. The construct has a selectable marker for the NPT?? gene. C and D: Southern blotting analysis of ex::phyA (C) and ex::appA (D) transgenic lines, using phyA and appA probes, respectively. Genomic DNA was digested by HindIII and EcoRI. C, M, Marker; WT, wild-type canola; 1–4,T3 transgenic plants of P3 line; 5–6, T3 transgenic plants of P11 line; D, 1–4, T3 transgenic plants of a18 line. E and F: Northern blotting analysis of phyA (E) and appA (F) expression in roots of transgenic lines using phyA and appA probe, respectively.