Figure 6.

Generation of the Δppabcg7 Knockout Mutant.

(A) Schematic diagram depicting the targeted replacement of Pp-ABCG7 with the nptII selectable marker by homologous recombination to yield a stable P. patens knockout mutant. (FR, flanking region).

(B) PCR screening of wild-type and Δppabcg7 genomic DNA with the PCR primer pairs shown in (A) showing the presence (Yes) or absence (No) of an amplified product. Primer pairs specific to Pp-ABCG7 generate a PCR product in the wild type but not in Δppabcg7.

(C) PCR screening of wild-type and Δppabcg7 cDNA with Pp-ABCG7 transcript-specific primers, showing that Pp-ABCG7 expression is abolished in Δppabcg7 (P, protonema; G, gametophore). Primers specific to P. patens actin (Pp-ACT2) were used as a control for cDNA quality.