Figure 2.

Genomic PCR, RT-PCR, and immunoblot analyses of ACT transformants. Eight-day-old suspension cells of wild-type (WT) Pn and ACT transgenic lines (15 and 22) cultured under PR conditions were analysed. For the PCR experiments (a,b), HvACT1 and HPT transgenes were amplified to verify their stable maintenance using genomic PCR (a) and transcription using RT-PCR (b) and actin was amplified as an endogenous standard. Full-length gels for genomic PCR and RT-PCR analyses are presented in Supplementary Fig. S5a,b, respectively. For the immunoblot analysis (c) the recombinant His-tag-free HvACT1 enzyme expressed in E. coli was used as a positive control (Ctrl) for the detection of the HvACT1 protein in the crude cell extracts. Asterisks indicate that the signals occurred by a non-specific cross-reaction of the anti-HvACT1 polyclonal antibody with some protein(s) in the crude extracts. Full-length blot is presented in Supplementary Fig. S5c.