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. 2015 Aug 26;6:657. doi: 10.3389/fpls.2015.00657

FIGURE 2.

FIGURE 2

Construction of transformants expressing sfGFP by using the promoters of the nitrate assimilation genes. (A) Schematic diagram showing the targeted insertion of sfGFP flanked with the NR, NIR, or NRT promoter into the chromosomal region upstream of the URA5.3 gene. The third to fifth lines indicate the expected genomic structures in which a single copy of the transformed sequence is inserted by double-crossover homologous recombination in the respective cases. The red asterisk shows the position of the point mutation in the URA5.3 gene in strain M4. The arrowheads indicate the positions of the PCR primers used in (B). (B) PCR analysis of independent transformants confirmed the homologous recombination events. The WT strain was used as a negative control. The genomic sequence of the WT strain is identical to that of M4 except for the point mutation in the URA5.3 gene in M4. The predicted size of the PCR product is 6.1-kb for NRp, NIRp, and NRTp or 4.2 kb for the WT, respectively. The positions and sequences of the primers No. 14 and No. 15 are shown in Supplementary Table S1.